Compound targeting IL-23A and TNF-alpha and uses thereof

ABSTRACT

The disclosure relates to compounds specific for IL23A and TNF-alpha, compositions comprising the compounds, and methods of use thereof. Nucleic acids, cells, and methods of production related to the compounds and compositions are also disclosed.

RELATED APPLICATION

This application is a divisional of U.S. application Ser. No.16/052,133, filed Aug. 1, 2018, now U.S. Pat. No. 10,793,629, which is adivisional of U.S. application Ser. No. 14/844,338 filed Sep. 3, 2015,now U.S. Pat. No. 10,059,763, and claims the benefit of the filing dateunder 35 U.S.C. § 119 of U.S. Provisional Application Ser. No.62/045,498, filed Sep. 3, 2014, and entitled Compound Targeting IL-23Aand TNF-ALPHA and Uses Thereof, the entire contents of each and all ofwhich are incorporated by reference herein.

SEQUENCE LISTING

The sequence listing in the file named “1157443o001003” having a size of534,059 that was created Aug. 26, 2020, is hereby incorporated byreference in its entirety.

BACKGROUND OF THE INVENTION

Inflammation involves an innate and adaptive immune response that isrequired for fighting infection. However, when the inflammation becomesunchecked autoimmune or autoinflammatory diseases, neurodegenerativedisease, and even cancer can develop. It is well established thatinhibiting activity of proinflammatory cytokines such as IL1, TNF-alpha,IL6, IL12, IL17, IL18, or IL23 reduces inflammation and suppressesspecific pathways that activate immune cells.

Interleukin 23 (IL23) is a heterodimeric cytokine consisting of twosubunits, p40 and p19. The p19 subunit is also referred to as IL-23A.While the p19 subunit is unique to IL23, the p40 subunit is shared withthe cytokine IL12. IL23 is emerging as a key regulator of pathogenicTh17, γδ T and innate lymphoid cells (ILCs) driving the production ofIL17, IL22 and other cytokines that lead to local tissue inflammationand damage. IL23 promotes upregulation of the matrix metalloproteaseMMP9, increases angiogenesis, reduces CD8+ T cell infiltration, and hasbeen implicated in the development of cancerous tumors. In addition, inconjunction with IL6 and TGF-beta1, IL23 stimulates naive CD4+ T cellsto differentiate into Th17 cells. In turn, the Th17 cells produce IL17,a proinflammatory cytokine that enhances T cell priming and stimulatesthe production of proinflammatory cytokines such as IL1, IL6, TNF-alpha,NOS-2, and also induces expression of chemokines resulting ininflammation and disease pathogenesis. IL23 exerts its effects via acell surface receptor composed of the IL12131 subunit of IL12 receptorpartnered with a unique IL23R subunit. Expression of the IL23R isrestricted to specific populations of immune cells and is foundprimarily on subsets of T cells (αβ and γδ TCR+) and NK cells.

In mice, genetic ablation of the IL23p19 gene results in selective lossof IL23 function accompanied by severely compromised T-dependent immuneresponses, including reduced production of antigen-specificimmunoglobulins and impaired delayed type hypersensitivity responses(Ghilardi N, et al. (2004) J. Immunol. 172(5): 2827-33). Knockout micedeficient in either IL23p40 or IL23p19, or in either subunit of the IL23receptor (IL23R and IL12-beta1), develop less severe symptoms in animalmodels of multiple sclerosis, arthritis and inflammatory bowel disease.Similar results have been obtained using an antibody specific forIL23p19 in EAE and a T cell mediated colitis model further substantiatesthe role of IL23 in these disease settings (Chen Y. et al. (2006) J.Clin. Invet. 116(5):1317-26; Elson C O. et al. (2007) Gastroenterology132(7): 2359-70). This highlights the importance of IL23 in chronicinflammation (Langowski et al. (2006) Nature 442 (7101): 461-5; Kikly K,et al. (2006) Curr. Opin. Immunol. 18 (6): 670-5). In addition, elevatedIL23 production has been implicated as being a major factor ininflammatory arthritis and in inflammatory autoimmune diseases(Adamopoulos et al. (2011) J. Immunol. 187: 593-594; and Langris et al.(2005) J. Exp. Med. 201:233-240). A connection between 1L23, itsdownstream cytokine 1L22, and bone formation has been published in amouse model system in which IL23 is overexpressed (Sherlock et al.(2012) Nat. Med. 18: 1069-76).

The homotrimeric TNF-α cytokine is expressed predominantly bymacrophages, lymphocytes, endothelial cells and fibroblasts and bindstwo distinct receptors: TNFRI, expressed on nearly all cell types andTNFRII, with more limited expression on immune cells (CD4+ T cells, NKcells). Like many TNF superfamily members, TNF-α exists as both membraneand soluble forms, the soluble form arising from cleavage of themembrane form by the ADAM12 metalloprotease (TACE, TNFα convertingenzyme). Both membrane-bound and soluble forms of the cytokine arebiologically active.

Tumor necrosis factor (TNF-alpha/TNF-α) is a proinflammatory cytokinethat stimulates the acute phase of inflammation. Tumor necrosis factorincreases vascular permeability through induction of IL8, therebyrecruiting macrophage and neutrophils to a site of infection. Oncepresent, activated macrophages continue to produce TNF-alpha, therebymaintaining and amplifying the inflammatory response. The primary roleof TNF-alpha is the regulation of immune cells; however, TNF-alpha isalso involved in the regulation of a wide spectrum of biologicalprocesses including cell proliferation, differentiation, apoptosis,lipid metabolism, and coagulation. TNF-alpha is able to induceinflammation, induce apoptotic cell death, inhibit tumorigenesis andinhibit viral replication.

Dysregulation of TNF-alpha production has been implicated in a varietyof human diseases, including autoimmune disease (e.g. rheumatoidarthritis (RA), Crohn's disease, multiple sclerosis), inflammatory boweldisease (IBD), ulcerative colitis, psoriasis, toxic shock, graft versushost disease, insulin resistance, Alzheimer's disease, cancer, and majordepression (Swardfager W, et al. (2010) Biol Psychiatry 68 (10):930-941; (Locksley R M, et al. (2001) Cell 104 (4): 487-501; Dowlati etal., (2010) Biol Psychiatry 67 (5): 446-457; Brynskov J. et al. (2002)Gut 51 (1): 37-43).

Antibodies have been used as biologic therapies for inhibition ofTNF-alpha and IL23 in order to treat a variety of inflammatory diseases.Infliximab (Centocor, Malvern, Pa.) described in U.S. Pat. Nos.6,277,969, 6,284,471, and 6,790,444, is a chimeric anti-TNF-alphamonoclonal IgG antibody bearing human IgG4 constant and mouse variableregions and is used clinically to treat rheumatoid arthritis, psoriaticarthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitisand plaque psoriasis. Monoclonal antibody adalimumab (clone D2E7; AbbottLaboratories, Abbott Park, Ill.) described in U.S. Pat. No. 6,090,382,is an anti-TNF-alpha therapy used clinically to treat rheumatoidarthritis, Crohn's disease, psoriasis, psoriatic arthritis, ankylosingspondylitis, and juvenile idiopathic arthritis. Golimumab is a TNF-alphablocker used to treat rheumatoid arthritis, psoriatic arthritis,ankylosing spondylitis, and ulcerative colitis. In addition, humanmonoclonal antibody ustekinumab (Centocor, Inc, Malvern, Pa.), describedin U.S. Pat. Nos. 6,902,734 and 7,166,285, is directed againstinterleukin 12 and interleukin 23 (specifically the p40 subunit), isclinically used to treat severe plaque psoriasis, and is further beinginvestigated for the treatment of psoriatic arthritis, multiplesclerosis, and sarcoidosis. However, anti-TNF-α therapies have reportedside effects [see for example: Keane J et al. (2001)]. Tuberculosis isassociated with infliximab, a tumor necrosis factor α-neutralizingagent. N Engl J Med 345 (15):1098-1104; Scheinfeld N. (2005) Adalimumab:a review of side effects. Expert Opin Drug Suf. 4(4):637-41;Chovel-Sella A et al. (2012) Clinical efficacy and adverse effects ofgolimumab in the treatment of rheumatoid arthritis. Isr Med Assoc J.14(6):390-4]. Identification of more efficacious treatments should allowfor administration of reduced dosages, as well as lower costs associatedwith the treatment.

There remains a need for compositions with increased efficacy fortreating and preventing autoimmune or inflammatory diseases.

SUMMARY OF THE INVENTION

Provided herein are compounds specific for TNF-alpha and IL23A,compositions comprising such compounds, as well as methods of use andproduction thereof.

Aspects of the disclosure relate to a compound comprising a firstpolypeptide and a second polypeptide, wherein:

(A) said first polypeptide comprises:

-   -   (i) a light chain variable domain of a first immunoglobulin        (VL1) specific for a first target protein;    -   (ii) a heavy chain variable domain of a second immunoglobulin        (VH2) specific for a second target protein; and    -   (iii) a hinge region, a heavy chain constant region 2 (CH2) and        a heavy chain constant region 3 (CH3); and

(B) said second polypeptide comprises:

-   -   (i) a light chain variable domain of the second immunoglobulin        (VL2) specific for said second target protein;    -   (ii) a heavy chain variable domain of the first immunoglobulin        (VH1) specific for said first target protein;

wherein:

-   -   (i) said VL1 and VH1 associate to form a binding site that binds        said first target protein;    -   (ii) said VL2 and VH2 associate to form a binding site that        binds said second target protein;    -   (iii) said heavy chain constant region 2 (CH2) comprises a        tyrosine at position 252, a threonine at position 254 and a        glutamic acid a position 256, numbered according to the EU index        as in Kabat for a conventional antibody; and    -   (iv) said first target protein is TNF-alpha and said second        target protein is IL-23A or said first target protein is IL-23A        and said second target protein is TNF-alpha,

wherein:

-   -   (i) said VL1 comprises SEQ ID NO:2, said VH1 comprises SEQ ID        NO:1, said VL2 comprises SEQ ID NO:8 and said VH2 comprises SEQ        ID NO:7; or    -   (ii) said VL1 comprises SEQ ID NO:4 or 6, said VH1 comprises SEQ        ID NO:3 or 5, said VL2 comprises SEQ ID NO:8 and said VH2        comprises SEQ ID NO:7; or    -   (iii) said VL1 comprises SEQ ID NO:8, said VH1 comprises SEQ ID        NO:7, said VL2 comprises SEQ ID NO:2 and said VH2 comprises SEQ        ID NO:1; or    -   (iv) said VL1 comprises SEQ ID NO:8, said VH1 comprises SEQ ID        NO:7, said VL2 comprises SEQ ID NO:4 or 6 and said VH2 comprises        SEQ ID NO:3 or 5.

In some embodiments, in (ii) said VL1 comprises SEQ ID NO:4, said VH1comprises SEQ ID NO:3, said VL2 comprises SEQ ID NO:8 and said VH2comprises SEQ ID NO:7. In some embodiments, in (ii) said VL1 comprisesSEQ ID NO:6, said VH1 comprises SEQ ID NO:5, said VL2 comprises SEQ IDNO:8 and said VH2 comprises SEQ ID NO:7. In some embodiments, in (iv)said VL2 comprises SEQ ID NO:4, said VH2 comprises SEQ ID NO:3, said VL1comprises SEQ ID NO:8 and said VH1 comprises SEQ ID NO:7. In someembodiments, in (iv) said VL2 comprises SEQ ID NO:6, said VH2 comprisesSEQ ID NO:5, said VL1 comprises SEQ ID NO:8 and said VH1 comprises SEQID NO:7.

In some embodiments, said first polypeptide further comprises a firstlinker between said VL1 and said VH2 and said second polypeptide furthercomprises a second linker between said VL2 and said VH1. In someembodiments, said first linker or said second linker comprises the aminoacid sequence GGGSGGG (SEQ ID NO:9). In some embodiments, said firstlinker and said second linker comprise the amino acid sequence GGGSGGG(SEQ ID NO:9).

In some embodiments, said first polypeptide further comprises a heavychain constant region 1 domain (CH1) and said second polypeptide furthercomprises a light chain constant region domain (CL), wherein said CL andsaid CH1 are associated together via a disulfide bond to form a C1domain.

In some embodiments, said first polypeptide further comprises a thirdlinker between said VH2 and said CH1 and said second polypeptide furthercomprises a fourth linker between said VH1 and said CL. In someembodiments, said third linker comprises the amino acid sequence FNRGES(SEQ ID NO:11). In some embodiments, said fourth linker comprises theamino acid sequence VEPKSS (SEQ ID NO:12). In some embodiments, saidthird linker comprises the amino acid sequence FNRGES (SEQ ID NO:11) andsaid fourth linker comprises the amino acid sequence VEPKSS (SEQ IDNO:12). In some embodiments, third linker or said fourth linkercomprises the amino acid sequence LGGGSG (SEQ ID NO:10). In someembodiments, said third linker and said fourth linker comprise the aminoacid sequence LGGGSG (SEQ ID NO:10).

In some embodiments, said heavy chain constant region 2 (CH2) comprisesan alanine at positions 234 and an alanine at position 235, numberedaccording to the EU index as in Kabat for a conventional antibody.

In some embodiments, the amino acid sequence of said hinge region, saidheavy chain constant region 2 (CH2) or said heavy chain constant region3 (CH3) is derived from a IgG1 or from a IgG4. In some embodiments, saidhinge region comprises the amino acid sequence EPKSCDKTHTCPPCP (SEQ IDNO:40).

In some embodiments, said compound comprises two said first polypeptidesand two said second polypeptides, wherein said two first polypeptidesarc associated together via at least one disulfide bond. In someembodiments, said compound comprises two said first polypeptides and twosaid second polypeptides, wherein said two first polypeptides areassociated together via at least one disulfide bond and wherein each ofsaid first polypeptide is associate to one said second polypeptide viaat least one disulfide bond.

In some embodiments,

-   -   (i) said first polypeptide comprises the amino acid sequence of        SEQ ID NO:13 and said second polypeptide comprises the amino        acid sequence of SEQ ID NO:14;    -   (ii) said first polypeptide comprises the amino acid sequence of        SEQ ID NO:15 and said second polypeptide comprises the amino        acid sequence of SEQ ID NO:16;    -   (iii) said first polypeptide comprises the amino acid sequence        of SEQ ID NO:17 and said second polypeptide comprises the amino        acid sequence of SEQ ID NO:18;    -   (iv) said first polypeptide comprises the amino acid sequence of        SEQ ID NO:19 and said second polypeptide comprises the amino        acid sequence of SEQ ID NO:20;    -   (v) said first polypeptide comprises the amino acid sequence of        SEQ ID NO:21 and said second polypeptide comprises the amino        acid sequence of SEQ ID NO:22;

(vi) said first polypeptide comprises the amino acid sequence of SEQ IDNO:23 and said second polypeptide comprises the amino acid sequence ofSEQ ID NO:24;

-   -   (vii) said first polypeptide comprises the amino acid sequence        of SEQ ID NO:25 and said second polypeptide comprises the amino        acid sequence of SEQ ID NO:26;    -   (viii) said first polypeptide comprises the amino acid sequence        of SEQ ID NO:27 and said second polypeptide comprises the amino        acid sequence of SEQ ID NO:28;    -   (ix) said first polypeptide comprises the amino acid sequence of        SEQ ID NO:29 and said second polypeptide comprises the amino        acid sequence of SEQ ID NO:30;    -   (x) said first polypeptide comprises the amino acid sequence of        SEQ ID NO:31 and said second polypeptide comprises the amino        acid sequence of SEQ ID NO:32;    -   (xi) said first polypeptide comprises the amino acid sequence of        SEQ ID NO:33 and said second polypeptide comprises the amino        acid sequence of SEQ ID NO:34; or    -   (xi) said first polypeptide comprises the amino acid sequence of        SEQ ID NO:35 and said second polypeptide comprises the amino        acid sequence of SEQ ID NO:36.

In some embodiments, wherein said compound comprises two said firstpolypeptides and two said second polypeptides, wherein said two firstpolypeptides are associated together via at least one disulfide bond.

In some embodiments, said compound comprises two said first polypeptidesand two said second polypeptides, and wherein the CH2 and CH3, and CH1if present, of one of the first polypeptides associates with the CH2 andCH3, and CH1 if present, of the other of the first polypeptides to forma tetravalent molecule. In some embodiments, said compound comprises twosaid first polypeptides and two said second polypeptides, wherein eachof said first polypeptides comprises a CH1, a CH2 and a CH3 and each ofsaid second polypeptides comprises a CL and wherein the CH2 and CH3 ofone of the first polypeptides associates with the CH2 and CH3 of theother of the first polypeptides and the CH1 of each said firstpolypeptides associates with the CL of one said second polypeptides toform a tetravalent molecule.

Other aspects of the disclosure relate to a first compound that competeswith a second compound for binding to IL-23A and to TNF-alpha, whereinsaid first compound comprises a third polypeptide and fourthpolypeptide, wherein:

(A) said third polypeptide comprises:

-   -   (i) a light chain variable domain of a first immunoglobulin        (VL1) specific for a first target protein;    -   (ii) a heavy chain variable domain of a second immunoglobulin        (VH2) specific for a second target protein; and    -   (iii) a hinge region, a heavy chain constant region 2 (CH2) and        a heavy chain constant region 3 (CH3); and

(B) said fourth polypeptide comprises:

-   -   (i) a light chain variable domain of the second immunoglobulin        (VL2) specific for said second target protein;    -   (ii) a heavy chain variable domain of the first immunoglobulin        (VH1) specific for said first target protein;

and wherein

-   -   (i) said VL1 and VH1 associate to form a binding site that binds        said first target protein;    -   (ii) said VL2 and VH2 associate to form a binding site that        binds said second target protein; and    -   (iii) said first target protein is TNF-alpha and said second        target protein is IL-23A or said first target protein is IL-23A        and said second target protein is TNF-alpha,

and wherein said second compound comprises a first polypeptide and asecond polypeptide, wherein:

-   -   (i) said first polypeptide comprises the amino acid sequence of        SEQ ID NO:13 and said second polypeptide comprises the amino        acid sequence of SEQ ID NO:14;    -   (ii) said first polypeptide comprises the amino acid sequence of        SEQ ID NO:15 and said second polypeptide comprises the amino        acid sequence of SEQ ID NO:16;    -   (iii) said first polypeptide comprises the amino acid sequence        of SEQ ID NO:17 and said second polypeptide comprises the amino        acid sequence of SEQ ID NO:18;    -   (iv) said first polypeptide comprises the amino acid sequence of        SEQ ID NO:19 and said second polypeptide comprises the amino        acid sequence of SEQ ID NO:20;    -   (v) said first polypeptide comprises the amino acid sequence of        SEQ ID NO:21 and said second polypeptide comprises the amino        acid sequence of SEQ ID NO:22;    -   (vi) said first polypeptide comprises the amino acid sequence of        SEQ ID NO:23 and said second polypeptide comprises the amino        acid sequence of SEQ ID NO:24;    -   (vii) said first polypeptide comprises the amino acid sequence        of SEQ ID NO:25 and said second polypeptide comprises the amino        acid sequence of SEQ ID NO:26;    -   (viii) said first polypeptide comprises the amino acid sequence        of SEQ ID NO:27 and said second polypeptide comprises the amino        acid sequence of SEQ ID NO:28;    -   (ix) said first polypeptide comprises the amino acid sequence of        SEQ ID NO:29 and said second polypeptide comprises the amino        acid sequence of SEQ ID NO:30;    -   (x) said first polypeptide comprises the amino acid sequence of        SEQ ID NO:31 and said second polypeptide comprises the amino        acid sequence of SEQ ID NO:32;    -   (xi) said first polypeptide comprises the amino acid sequence of        SEQ ID NO:33 and said second polypeptide comprises the amino        acid sequence of SEQ ID NO:34; or    -   (xii) said first polypeptide comprises the amino acid sequence        of SEQ ID NO:35 and said second polypeptide comprises the amino        acid sequence of SEQ ID NO:36.

Yet other aspects of the disclosure relate to a pharmaceuticalcomposition comprising a compound described herein, such as a compounddescribed above.

Other aspects of the disclosure relate to a method of treating anautoimmune or an inflammatory disease comprising administering acompound described herein, such as a compound described above, or apharmaceutical composition comprising said compound to a subject.

Yet other aspects of the disclosure relate to a compound describedherein, such as a compound described above, for use in medicine. In someembodiments, said use is the treatment of an autoimmune or aninflammatory disease.

Other aspects of the disclosure relate to a pharmaceutical compositioncomprising a compound described herein, such as a compound describedabove, for use in medicine. In some embodiments, said use is thetreatment of an autoimmune or an inflammatory disease.

Yet other aspects of the disclosure relate to a use of a compounddescribed herein, such as a compound described above, in the manufactureof a medicament for use in medicine. In some embodiments, said use isthe treatment of an autoimmune or an inflammatory disease.

Other aspects of the disclosure relate to a use of a pharmaceuticalcomposition described herein, such as a pharmaceutical compositiondescribed above, in the manufacture of a medicament for use in medicine.In some embodiments, said use is the treatment of an autoimmune or aninflammatory disease.

Yet other aspects of the disclosure relate to a nucleic acid comprisinga nucleotide sequence encoding a polypeptide described herein, such as apolypeptide described above. Other aspects of the disclosure relate to avector comprising said nucleic acid. In some embodiments, the vectorcomprises a promoter operably linked to said nucleic acid. Other aspectsof the disclosure relate to a cell comprising said nucleic acid or saidvector.

Other aspects of the disclosure relate to a method of producing acompound or polypeptide as described herein, such as a polypeptidedescribed above, comprising obtaining a cell described herein, such acell described above, and expressing a nucleic acid as described hereinin said cell. In some embodiments, the method further comprisesisolating and purifying said polypeptide or compound.

The details of one or more embodiments of the disclosure are set forthin the description below. Other features or advantages of the presentdisclosure will be apparent from the following drawings and detaileddescription of several embodiments, and also from the appending claims.

BRIEF DESCRIPTION OF THE DRAWINGS

The following drawings form part of the present specification and areincluded to further demonstrate certain aspects of the presentdisclosure, which can be better understood by reference to one or moreof these drawings in combination with the detailed description ofspecific embodiments presented herein.

FIG. 1A is a diagram of an exemplary compound specific for TNF-alpha andIL23A. The first polypeptide chain contains CH3, CH2, VH₂ (VH_(II)) andVL₁ (VL_(I)) domains. The second polypeptide chain contains VH₁ (VH_(I))and VL₂(VL_(II)) domains. VL₁ and VH₁ are specific for a first targetprotein (either TNF-alpha or IL23A) and VL₂ and VH₂ are specific for asecond target protein (either IL23A or TNF-alpha). The upper panel showseach polypeptide chain separately. The lower panel shows a tetravalentcompound formed through association of the CH2 and CH3 domains of onefirst polypeptide with the CH2 and CH3 domains of another firstpolypeptide. The binding domains for the first and second target proteinare formed through association of VH₁ and VL₁ and through associationVH₂ and VL₂, respectively.

FIG. 1B is a diagram of another exemplary compound specific forTNF-alpha and IL23A. The first polypeptide chain contains CH3, CH2, CH1,VH₂ (VH_(II)) and VL₁ (VL_(I)) domains. The second polypeptide chaincontains CL, VH₁ (VH_(I)) and VL₂ (VL_(II)) domains. VL_(I) and VH₁ arespecific for a first target protein (either TNF-alpha or IL23A) and VL₂and VH₂ are specific for a second target protein (either IL23A orTNF-alpha). The upper panel shows each polypeptide chain separately. Thelower panel shows a tetravalent compound formed through association ofthe CH2 and CH3 domains of one first polypeptide with the CH2 and CH3domains of another first polypeptide. The binding domains for the firstand second target protein are formed through association of VH₁ and VL₁and through association VH₂ and VL₂, respectively. The compound isfurther associated through interactions between the CL and CH1 domains.

FIG. 2 is a graph showing the serum concentrations of Compound M and itsYTE mutant Compound A in male cynomolgus monkey (mean±SD; N=3) after 1mg/kg IV 10 minute infusion.

FIG. 3 is a graph show serum concentrations of Compound 0 and itscorresponding YTE mutant Compound E in male cynomolgus monkey (mean±SD;N=3) after 1 mg/kg IV 10 minute infusion.

FIG. 4 is a series of graphs and a table showing that compound Emaintained functional potency vs. IL23 in vivo. Mice were dosedequimolar with either control antibody 3 (IL23A monoclonal antibody) orcompound E and challenged with human IL23 twice to induce earinflammation. Twenty four hours after the final injection, ears werecollected and analyzed for mouse IL17A and mouse IL22 as a measure offunctional blockade of IL23. Compound E maintained functional potency invivo vs. control antibody 3 (IL23A monoclonal antibody) based onterminal exposure and level of efficacy. Control antibody 3: opensquares, triangles and diamonds. Compound E: full squares, triangles anddiamonds. MW: molecular weight.

FIG. 5 is a series of graphs and a table showing that Compound Emaintained functional potency vs. TNF in vivo. Mice were dosed equimolarwith either control antibody 2 (TNFα monoclonal antibody) or compound Eand challenged with human TNF. Two hours after the challenge, wholeblood was collected and serum analyzed for mouse KC and mouse IL-6 as ameasure of functional blockade of TNF. Compound E maintained functionalpotency in vivo vs. anti-TNF based on terminal exposure and level ofefficacy. Control antibody 3: open squares, triangles and diamonds.Compound E: full squares, triangles and diamonds. MW: molecular weight.

DETAILED DESCRIPTION OF THE INVENTION

Described herein compounds that bind to both TNF-alpha (also referred toherein as TNF-α or TNFa) and IL23A (also referred to as IL23p19 orIL-23A). To date, there have been no approved compounds that target bothTNF-alpha and IL23A. There are limited studies with simultaneousneutralization of two/more key inflammatory mediators usingbio-therapeutics approach. While these studies failed to showimprovement in clinical outcomes that were measured for rheumatoidarthritis (RA), a bi-functional therapeutic targeting the samecombination has not been described to date. In addition, suchcombinations may increase side effects, such as the risk of infection(see, e.g., Genovese, M. C., Cohen, S., Moreland, L., Lium, D., Robbins,S., et al. (2004). Arth. Rheum. 50, 1412-9; Genovese, M. C., Cohen, S.,Moreland, L., Lium, D., Robbins, S., et al. (2004). Arth. Rheum. 50,1412-9; and Weinblatt, M., Schiff, M., Goldman, A. Kremer, J., Luggen,M., et al. (2007). Ann. Rheum. Dis.66, 228-34). Further, suchbi-specific compounds have been difficult to design, due to issuesrelated to solubility (e.g., aggregation) and stability (e.g., poorpharmacokinetics).

Surprisingly, the compounds described herein that bind to both TNF-alphaand IL23A have been found to have similar or improved propertiescompared to individual antibodies that target either IL23A or TNF-alpha.These compounds were also found to have suitable pharmacokinetics andwere soluble at suitable ranges for dosing purposes. Further, in someembodiments, there are advantages of single administration over multipleindividual dose administration from the perspective of side effects ofthe individual therapies, and lower dosage. In addition, in someembodiments, the CMC properties of the compounds showed that compoundshad low aggregation. In one aspect, exemplary compounds showedparticularly low aggregation. It was also shown that the linkers wereoptimized to improve stability and prevented cleavage and that the YTEmutation improved Fc Rn affinity. The compounds described herein arebelieved to have one or more advantageous properties, e.g., decreasedside effects, increased ease and safety of administration, an increasedhalf-life, increased binding affinity, or increased inhibitory activity,compared to standard antibody molecules, e.g., an IgG molecule orantigen-binding fragment (Fab).

Accordingly, aspects of the disclosure relate to compounds specific forboth TNF-alpha and IL23A, as well as methods of use and production ofsuch compounds.

Compounds

Aspects of the disclosure relate to a compound specific for bothTNF-alpha and IL23A. An exemplary protein sequence for TNF-alpha and anexemplary protein sequence for IL23A are shown below.

>NP_000585.2 - TNF-alpha [Homo sapiens] (SEQ ID NO: 144)MSTESMIRDVELAEEALPKKTGGPQGSRRCLFLSLFSFLIVAGATTLFCLLHFGVIGPQREEFPRDLSLISPLAQAVRSSSRTPSDKPVAHVVANPQAEGQLQWLNRRANALLANGVELRDNQLVVPSEGLYLIYSQVLFKGQGCPSTHVLLTHTISRIAVSYQTKVNLLSAIKSPCQRETPEGAEAKPWYEPIYLGGVFQLEKGDRLSAEINRPDYLDFAESGQVYFGIIAL >NP_057668.1 - IL23A [Homo sapiens](SEQ ID NO: 145) MLGSRAVMLLLLLPWTAQGRAVPGGSSPAWTQCQQLSQKLCTLAWSAHPLVGHMDLREEGDEETTNDVPHIQCGDGCDPQGLRDNSQFCLQRIHQGLIFYEKLLGSDIFTGEPSLLPDSPVGQLHASLLGLSQLLQPEGHHWETQQIPSLSPSQPWQRLLLRFKILRSLQAFVAVAARVFAHGAATLSP (amino acids1-19 are a predicted signal sequence)

In some embodiments, the compound comprises a first polypeptide and asecond polypeptide. In some embodiments, the first polypeptide comprises(i) a light chain variable domain of a first immunoglobulin (VL1)specific for a first target protein, (ii) a heavy chain variable domainof a second immunoglobulin (VH2) specific for a second target protein;and (iii) a hinge region, a heavy chain constant region 2 (CH2) and aheavy chain constant region 3 (CH3). In some embodiments, the firstpolypeptide further comprises a heavy chain constant region 1 (CH1). Insome embodiments, the second polypeptide comprises: (i) a light chainvariable domain of the second immunoglobulin (VL2) specific for thesecond target protein; (ii) a heavy chain variable domain of the firstimmunoglobulin (VH1) specific for the first target protein. In someembodiments, the first polypeptide further comprises a light chainconstant region (CL).

It is to be understood that the variable domains and constantdomains/regions of the first polypeptide can be in any order and thatthe variable domains and constant domains/regions (if any) of the secondpolypeptide can be in any order. Multiple exemplary configurations forthe domains/regions on the first and second polypeptide from N-terminusto C-terminus are shown below.

First polypeptide configuration 1: N-VL1-VH2-hinge-CH2-CH3-C

First polypeptide configuration 2: N-VH2-VL1-hinge-CH2-CH3-C

First polypeptide configuration 3: N-VL1-VH2-CH1-hinge-CH2-CH3-C

First polypeptide configuration 4: N-VH2-VL1-CH1-hinge-CH2-CH3-C

Second polypeptide configuration 1: N-VL2-VH1-C

Second polypeptide configuration 2: N-VH1-VL2-C

Second polypeptide configuration 3: N-VL2-VH1-CL-C

Second polypeptide configuration 4: N-VH1-VL2-CL-C

Exemplary configurations of the compound arc shown in FIGS. 1A and 1B.In some embodiments, the compound comprises the first polypeptide inconfiguration 1 and the second polypeptide in configuration 1. In someembodiments, the compound comprises the first polypeptide inconfiguration 3 and the second polypeptide in configuration 3.

In some embodiments, the variable regions of the first polypeptide andthe second polypeptide associate with one another to form a binding sitefor the first target protein and a binding site for the second targetprotein. In some embodiments, the VL1 of the first polypeptide and theVH1 of the second polypeptide associate to form a binding site thatbinds the first target protein and the VL2 of the second polypeptide andthe VH2 of the first polypeptide associate to form a binding site thatbinds the second target protein. In some embodiments, the first targetprotein is TNF-alpha and the second target protein is IL23A. In otherembodiments, the first target protein is IL23A and the second targetprotein is TNF-alpha. It is to be understood that the terms “first” and“second” are not meant to imply a level of importance to either targetprotein.

Exemplary combinations of sequences for each of VL1, VH1, VL2, and VH2are provided below in Table 1 and also in Table 2A in Example 1.

TABLE 1 Exemplary combinations of sequences for VL1, VH1, VL2, and VH2.Combination Number VL1 sequence VH1 sequence VL2 sequence VH2 sequence 1SEQ ID NO: 2 SEQ ID NO: 1 SEQ ID NO: 8 SEQ ID NO: 7 2 SEQ ID NO: 8 SEQID NO: 7 SEQ ID NO: 2 SEQ ID NO: 1 3 SEQ ID NO: 8 SEQ ID NO: 7 SEQ IDNO: 4 SEQ ID NO: 3 4 SEQ ID NO: 8 SEQ ID NO: 7 SEQ ID NO: 6 SEQ ID NO: 55 SEQ ID NO: 8 SEQ ID NO: 7 SEQ ID NO: 4 SEQ ID NO: 5 6 SEQ ID NO: 8 SEQID NO: 7 SEQ ID NO: 6 SEQ ID NO: 3 7 SEQ ID NO: 4 SEQ ID NO: 3 SEQ IDNO: 8 SEQ ID NO: 7 8 SEQ ID NO: 4 SEQ ID NO: 5 SEQ ID NO: 8 SEQ ID NO: 79 SEQ ID NO: 6 SEQ ID NO: 3 SEQ ID NO: 8 SEQ ID NO: 7 10 SEQ ID NO: 6SEQ ID NO: 5 SEQ ID NO: 8 SEQ ID NO: 7

In some embodiments, the compound comprises a VL1 sequence comprising afirst light chain CDR1, CDR2, and CDR3 and a VH1 sequence comprising afirst heavy chain CDR1, CDR2, and CDR3, a VL2 sequence comprising asecond light chain CDR1, CDR2 and CDR3, and a VH2 sequence comprising asecond heavy chain CDR1, CDR2, and CDR3. In some embodiments, the CDRsare the CDRs of one or more VL1, VH1, VL2, and VH2 sequences provided inTable 1 or Table 2A. Exemplary light chain and heavy chain CDR sequencesfor the VL1, VH1, VL2, and VH2 sequences provided in Table 1 arc shownbelow:

SEQ ID NO: 1 CDRs: (SEQ ID NO: 146) DYAMH (CDR1), (SEQ ID NO: 147)AITWNSGHIDYADSVEG (CDR2), (SEQ ID NO: 148) VSYLSTASSLDY (CDR3)SEQ ID NO: 2 CDRs: (SEQ ID NO: 149) RASQGIRNYLA (CDR1), (SEQ ID NO: 150)AASTLQS (CDR2), (SEQ ID NO: 151) QRYNRAPYT (CDR3)SEQ ID NO: 3 and SEQ ID NO: 5 CDRs: (SEQ ID NO: 152) SYAMH (CDR1),(SEQ ID NO: 153) FMSYDGSNKKYADSVKG (CDR2), (SEQ ID NO: 154)NYYYYGMDV (CDR3) SEQ ID NO: 4 and SEQ ID NO: 6 CDRs: (SEQ ID NO: 155)RASQSVYSYLA (CDR1), (SEQ ID NO: 156) DASNRAT (CDR2), (SEQ ID NO: 157)QQRSNWPPFT (CDR3) SEQ ID NO: 7 CDRs: (SEQ ID NO: 158) DQTIH (CDR1),(SEQ ID NO: 159) YIYPRDDSPKYNENFKG (CDR2), (SEQ ID NO: 160)PDRSGYAWFIY (CDR3) SEQ ID NO: 8 CDRs: (SEQ ID NO: 161)KASRDVAIAVA (CDR1), (SEQ ID NO: 162) WASTRHT (CDR2), (SEQ ID NO: 163)HQYSSYPFT (CDR3)

In some embodiments, the compound comprises a VH1, VL1, VH2, and/or VL2that comprises a sequence that is at least 80% (e.g., 85%, 90%, 95%,96%, 97%, 98%, or 99%) identical to a sequence described in Table 1. The“percent identity” of two amino acid sequences is determined using thealgorithm of Karlin and Altschul Proc. Natl. Acad. Sci. USA 87:2264-68,1990, modified as in Karlin and Altschul Proc. Natl. Acad. Sci. USA90:5873-77, 1993. Such an algorithm is incorporated into the NBLAST andXBLAST programs (version 2.0) of Altschul, et al. J. Mol. Biol.215:403-10, 1990. BLAST protein searches can be performed with theXBLAST program, score=50, wordlength=3 to obtain amino acid sequenceshomologous to the protein molecules of interest. Where gaps existbetween two sequences, Gapped BLAST can be utilized as described inAltschul et al., Nucleic Acids Res. 25(17):3389-3402, 1997. Whenutilizing BLAST and Gapped BLAST programs, the default parameters of therespective programs (e.g., XBLAST and NBLAST) can be used.

In some embodiments, the compound comprises a VH1, VL1, VH2, and/or VL2that comprises a sequence comprising one or more (e.g., 1, 2, 3, 4, 5,6, 7, 8, 9, 10 or more) mutations in a sequence described in Table 1.Such mutations can be conservative amino acid substitutions. As usedherein, a “conservative amino acid substitution” refers to an amino acidsubstitution that does not alter the relative charge or sizecharacteristics of the protein in which the amino acid substitution ismade. Conservative substitutions of amino acids include, for example,substitutions made amongst amino acids within the following groups: (a)M, I, L, V; (b) F, Y, W; (c) K, R, H; (d) A, G; (e) S, T; (f) Q, N; and(g) E, D.

The amino acid sequences of the hinge region, CH2 and CH3 of thecompound (and optionally the CH1 and CL, if the compound contains suchregions) may be derived from any appropriate source, e.g., a constantregion of an antibody such as an IgG1, IgG2, IgG3, or IgG4. Antibodyheavy and light chain constant regions amino acid sequences are wellknown in the art, e.g., those provided in the IMGT database(www.imgt.org) or at www.vbase2.org/vbstat.php, both of which areincorporated by reference herein. In some embodiments, the amino acidsequences of the CH2 and CH3 are derived from an IgG1 or an IgG4 (e.g.,SEQ ID NO: 39 or 37). In some embodiments, the CL comprises the aminoacid sequence of a kappa CL or a lambda CL. In some embodiments, thehinge region comprises the amino acid sequence EPKSCDKTHTCPPCP (SEQ IDNO:40).

In some embodiments, the CH2 and/or CH3 of the compound (and optionallythe CH1 and CL, if the compound contains such regions) may comprise oneor more amino acid substitutions that differ from a wild type CH2 orCH3, e.g., one or more amino acid substitutions in a wild type IgG1 CH2or CH3 or one or more amino acid substitutions in a wild type IgG4 CH2or CH3 (SEQ ID NO: 39 provides an exemplary wild-type IgG1). Suchsubstitutions are known in the art (see, e.g., U.S. Pat. Nos. 7,704,497,7,083,784, 6,821,505, 8,323,962, 6,737,056, and 7,416,727).

In some embodiments, the CH2 comprises an amino acid substitution at234, 235, 252, 254, and/or 256, numbered according to the EU index as inKabat for a conventional antibody (Kabat et al. Sequences of Proteins ofImmunological Interest, U.S. Department of Health and Human Services,1991, which is incorporated by reference herein in its entirety). It isto be understood that all amino acid positions described herein refer tothe numbering of the EU index as in Kabat for a conventional antibody.In some embodiments, the CH2 comprises an amino acid substitution atposition 252, 254, and/or 256. In some embodiments, the amino acid atposition 252 is tyrosine, phenylalanine, serine, tryptophan, orthreonine. In some embodiments, the amino acid at position 254 isthreonine. In some embodiments, the amino acid at position 254 isserine, arginine, glutamine, glutamic acid, or aspartic acid. In someembodiments, the CH2 comprises a tyrosine at position 252, a threonineat position 254 and a glutamic acid a position 256 (referred to hereinas a YTE mutant). In some embodiments, the CH2 comprises an amino acidsubstitution at position 234 and/or 235. In some embodiments, the CH2comprises an alanine at position 234 and an alanine at position 235,also referred to herein as KO mutant. In some embodiments, the CH2comprises a tyrosine at position 252, a threonine at position 254, aglutamic acid a position 256, an alanine at position 234 and an alanineat position 235, also referred to herein as KO-YTE mutant.

In some embodiments, one or more linkers may be used to connectdomains/regions together on the first and/or second polypeptide. Forexample, the first polypeptide may comprise a linker between the VL1 andVH2. If the first polypeptide comprises a CH1, the first polypeptide maycomprise a linker between the VL1 or VH2 (depending on the configurationdiscussed above) and the CH1 (e.g., VL1-linker-CH1 or VH2-linker-CH1).In another example, the second polypeptide may comprise a linker betweenthe VL2 and VH1. If the second polypeptide further comprises a CL, thesecond polypeptide may further comprise a linker between the VL2 or VH1(depending on the configuration discussed above) and the CL (e.g.,VL2-linker-CL or VH1-linker-CL). It is to be understood that any numberof linkers may be used to connect any domain or region to any otheranother domain or region on the first polypeptide and/or that any numberof linkers may be used to connect any domain or region to any otheranother domain or region on the second polypeptide.

Any suitable linker known in the art is contemplated for use herein. Insome embodiments, the linker is a peptide linker. In some embodiments,the peptide linker comprises at least two amino acids, e.g., 2, 3, 4, 5,6, 7, 8, 9, 10 or more amino acids. In some embodiments, the peptidelinker is no more than 50, 40, 30, 25, 20, 19, 18, 17, 16, 15, 14, 13,12, 11, 10, 9, 8, 7, 6, 5, 4, 3, or 2 amino acids in length. In someembodiments, the peptide linker is between 2 and 50, 2 and 40, 2 and 30,2 and 20, 2 and 10, 2 and 9, 2 and 8, 2 and 7, or 2 and 6 amino acids inlength. In some embodiments, the peptide linker comprises the amino acidsequence GGGSGGG (SEQ ID NO:9), LGGGSG (SEQ ID NO:10), FNRGES (SEQ IDNO:11), VEPKSS (SEQ ID NO:12), or a combination thereof. In someembodiments, the peptide linker may comprise multiple copies of a linkersequence, e.g., multiple copies of the sequence GGGSGGG (SEQ ID NO:9),LGGGSG (SEQ ID NO:10), FNRGES (SEQ ID NO:11), VEPKSS (SEQ ID NO:12), ora combination thereof.

In some embodiments, the compound comprises two first polypeptides andtwo second polypeptides. In some embodiments, the CH2 and CH3 of one ofthe first polypeptides associates with the CH2 and CH3 of the other ofthe first polypeptides to form a tetravalent molecule (e.g., the twofirst polypeptides dimerize through associations between theirrespective CH2 and CH3 domains to form a tetravalent molecule comprisingtwo binding sites specific for the first target protein and two bindingsites specific for the second target protein). If the first polypeptidefurther comprises a CH1 domain, the CH1 domain may also participate information of a tetravalent molecule (e.g., the two first polypeptidesdimerize through associations between their respective CH1, CH2 and CH3domains to form a tetravalent molecule comprising two binding sites forthe first target protein and two binding sites for the second targetprotein). In some embodiments, the two first polypeptides are associatedtogether via at least one disulfide bond.

Also contemplated herein are other compounds that compete for bindingwith a compound as described herein, e.g., a test compound that competeswith a compound as described herein for binding to TNF-alpha and IL23A.In some embodiments, the test compound may have at least 80%, 85%, 90%,95%, 96%, 97%, 98%, or 99% sequence identity with a compound asdescribed herein. Competitive binding may be determined using any assayknown in the art, e.g., equilibrium binding, ELISA, surface plasmonresonance, or spectroscopy.

In some embodiments, the compound described herein specifically binds toboth TNF-alpha and IL23A. A compound that “specifically binds” to anantigen or an epitope is a term well understood in the art, and methodsto determine such specific binding arc also well known in the art. Amolecule is said to exhibit “specific binding” if it reacts orassociates more frequently, more rapidly, with greater duration and/orwith greater affinity with a particular target antigen than it does withalternative targets. A compound “specifically binds” to a target antigenor epitope if it binds with greater affinity, avidity, more readily,and/or with greater duration than it binds to other substances. Forexample, a compound that specifically (or preferentially) binds to anantigen (e.g., TNF-alpha or IL23A) or an antigenic epitope therein is acompound that binds this target antigen with greater affinity, avidity,more readily, and/or with greater duration than it binds to otherantigens or other epitopes in the same antigen. It is also understood byreading this definition that, for example, a compound that specificallybinds to a first target antigen may or may not specifically orpreferentially bind to a second target antigen. As such, “specificbinding” or “preferential binding” does not necessarily require(although it can include) exclusive binding. Generally, but notnecessarily, reference to binding means preferential binding. In someexamples, a compound that “specifically binds” to a target antigen or anepitope thereof may not bind to other antigens or other epitopes in thesame antigen.

In some embodiments, a compound as described herein has a suitablebinding affinity for TNF-alpha and IL23 or antigenic epitopes thereof.As used herein, “binding affinity” refers to the apparent associationconstant or K_(A). The K_(A) is the reciprocal of the dissociationconstant (K_(D)). The compound described herein may have a bindingaffinity (K_(D)) of at least 10⁻⁵, 10⁻⁶, 10⁻⁷, 10⁻⁸, 10⁻⁹, 10⁻¹⁰, 10⁻¹¹,10⁻¹² M or lower for one or both of the target antigens or antigenicepitopes. An increased binding affinity corresponds to a decreasedK_(D). In some embodiments, the compound described herein has a bindingaffinity (K_(D)) of at least 10⁻¹¹M or lower for one or both of thetarget antigens or antigenic epitopes. Higher affinity binding of acompound for a first antigen and a second antigen relative to a thirdantigen can be indicated by a higher K_(A) (or a smaller numerical valueK_(D)) for binding the first antigen and second antigen than the K_(A)(or numerical value K_(D)) for binding the third antigen. In such cases,the compound has specificity for the first antigen and second antigen(e.g., a first protein in a first conformation or mimic thereof and asecond protein in a first conformation or mimic thereof) relative to thethird antigen (e.g., the same first or second protein in a secondconformation or mimic thereof; or a third protein). Differences inbinding affinity (e.g., for specificity or other comparisons) can be atleast 1.5, 2, 3, 4, 5, 10, 15, 20, 37.5, 50, 70, 80, 91, 100, 500, 1000,10,000 or 10⁵ fold.

Binding affinity (or binding specificity) can be determined by a varietyof methods including, equilibrium binding, ELISA, surface plasmonresonance, or spectroscopy (e.g., using a fluorescence assay). Exemplaryconditions for evaluating binding affinity are in HBS-P buffer (10 mMHEPES pH7.4, 150 mM NaCl, 0.005% (v/v) Surfactant P20). These techniquescan be used to measure the concentration of bound binding protein as afunction of target protein concentration. The concentration of boundbinding protein ([Bound]) is related to the concentration of free targetprotein ([Free]) and the concentration of binding sites for the bindingprotein on the target where (N) is the number of binding sites pertarget molecule by the following equation:[Bound]=[N][Free]/(Kd+[Free])

It is not always necessary to make an exact determination of K_(A),though, since sometimes it is sufficient to obtain a quantitativemeasurement of affinity, e.g., determined using a method such as ELISAor FACS analysis, is proportional to K_(A), and thus can be used forcomparisons, such as determining whether a higher affinity is, e.g.,2-fold higher, to obtain a qualitative measurement of affinity, or toobtain an inference of affinity, e.g., by activity in a functionalassay, e.g., an in vitro or in vivo assay.

In some embodiments, the compound comprises a first polypeptide and asecond polypeptide as defined in Table 2A. In some embodiments, thecompound comprises:

-   -   (i) a first polypeptide comprises the amino acid sequence of SEQ        ID NO:13 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:14;    -   (ii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:15 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:16;    -   (iii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:17 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:18;    -   (iv) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:19 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:20;    -   (v) a first polypeptide comprises the amino acid sequence of SEQ        ID NO:21 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:22;    -   (vi) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:23 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:24;    -   (vii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:25 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:26;    -   (viii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:27 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:28;    -   (ix) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:29 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:30;    -   (x) a first polypeptide comprises the amino acid sequence of SEQ        ID NO:31 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:32;    -   (xi) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:33 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:34;    -   (xii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:35 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:36;    -   (xiii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:44 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:45;    -   (xiv) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:46 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:47;    -   (xv) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:48 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:49;    -   (xvi) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:50 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:51;    -   (xvii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:52 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:53;    -   (xviii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:54 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:55;    -   (xix) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:56 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:57;    -   (xx) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:58 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:59;    -   (xxi) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:60 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:61;    -   (xxii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:62 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:63;    -   (xxiii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:64 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:65;    -   (xxiv) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:66 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:67;    -   (xxv) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:68 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:69;    -   (xxvi) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:70 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:71;    -   (xxvii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:72 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:73;    -   (xxviii) a first polypeptide comprises the amino acid sequence        of SEQ ID NO:74 and a second polypeptide comprises the amino        acid sequence of SEQ ID NO:75;    -   (xxix) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:76 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:77;    -   (xxx) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:78 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:79;    -   (xxxi) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:80 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:81;    -   (xxxii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:82 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:83;    -   (xxxiii) a first polypeptide comprises the amino acid sequence        of SEQ ID NO:84 and a second polypeptide comprises the amino        acid sequence of SEQ ID NO:85;    -   (xxxiv) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:86 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:87;    -   (xxxv) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:88 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:89;    -   (xxxvi) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:90 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:91;    -   (xxxvii) a first polypeptide comprises the amino acid sequence        of SEQ ID NO:92 and a second polypeptide comprises the amino        acid sequence of SEQ ID NO:93;    -   (xxxviii) a first polypeptide comprises the amino acid sequence        of SEQ ID NO:94 and a second polypeptide comprises the amino        acid sequence of SEQ ID NO:95;    -   (xxxix) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:96 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:97;    -   (xl) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:98 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:99;    -   (xli) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:100 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:101;    -   (xlii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:102 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:103;    -   (xliii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:104 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:105;    -   (xliv) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:106 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:107;    -   (xlv) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:108 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:109;    -   (xlvi) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:110 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:111;    -   (xlvii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:112 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:113;    -   (xlviii) a first polypeptide comprises the amino acid sequence        of SEQ ID NO:114 and a second polypeptide comprises the amino        acid sequence of SEQ ID NO:115;    -   (xlix) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:116 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:117;    -   (l) a first polypeptide comprises the amino acid sequence of SEQ        ID NO:118 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:119;    -   (li) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:120 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:121;    -   (lii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:122 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:123;    -   (liii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:124 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:125;    -   (liv) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:126 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:127;    -   (lv) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:128 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:129;    -   (lvi) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:130 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:131;    -   (lvii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:132 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:133;    -   (lviii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:134 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:135;    -   (lix) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:136 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:137;    -   (lx) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:138 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:139;    -   (lxi) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:140 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:141; or    -   (lxii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:142 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:143.

In some embodiments, the compound comprises:

-   -   (i) a first polypeptide comprises the amino acid sequence of SEQ        ID NO:13 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:14;    -   (ii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:15 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:16;    -   (iii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:17 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:18;    -   (iv) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:19 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:20;    -   (v) a first polypeptide comprises the amino acid sequence of SEQ        ID NO:21 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:22;    -   (vi) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:23 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:24;    -   (vii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:25 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:26;    -   (viii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:27 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:28;    -   (ix) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:29 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:30;    -   (x) a first polypeptide comprises the amino acid sequence of SEQ        ID NO:31 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:32;    -   (xi) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:33 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:34; or    -   (xii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:35 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:36.        Methods of Producing Compounds, Nucleic Acids, Vectors, and        Cells

Aspects of the disclosure also include nucleic acids that encodecompounds described herein or polypeptides described herein (e.g., firstor second polypeptides described herein), which may be encoded togetheror separately. The polynucleotides encoding the compounds describedherein or polypeptides described herein may be obtained, and thenucleotide sequence of the polynucleotides determined, by any methodknown in the art.

In some embodiments, the nucleic acid is comprised within a vector, suchas an expression vector. In some embodiments, the vector comprises apromoter operably linked to the nucleic acid.

A variety of promoters can be used for expression of the compoundsdescribed herein or polypeptides described herein, including, but notlimited to, cytomegalovirus (CMV) intermediate early promoter, a viralLTR such as the Rous sarcoma virus LTR, HIV-LTR, HTLV-1 LTR, the simianvirus 40 (SV40) early promoter, E. coli lac UV5 promoter, and the herpessimplex tk virus promoter.

Regulatable promoters can also be used. Such regulatable promotersinclude those using the lac repressor from E. coli as a transcriptionmodulator to regulate transcription from lac operator-bearing mammaliancell promoters [Brown, M. et al., Cell, 49:603-612 (1987)], those usingthe tetracycline repressor (tetR) [Gossen, M., and Bujard, H., Proc.Natl. Acad. Sci. USA 89:5547-5551 (1992); Yao, F. et al., Human GeneTherapy, 9:1939-1950 (1998); Shockelt, P., et al., Proc. Natl. Acad.Sci. USA, 92:6522-6526 (1995)]. Other systems include FK506 dimer, VP16or p65 using astradiol, RU486, diphenol murislerone, or rapamycin.Inducible systems are available from Invitrogen, Clontech and Ariad.

Regulatable promoters that include a repressor with the operon can beused. In one embodiment, the lac repressor from Escherichia coli canfunction as a transcriptional modulator to regulate transcription fromlac operator-bearing mammalian cell promoters [M. Brown et al., Cell,49:603-612 (1987)]; Gossen and Bujard (1992); [M. Gossen et al., Natl.Acad. Sci. USA, 89:5547-5551 (1992)] combined the tetracycline repressor(tetR) with the transcription activator (VP 16) to create atetR-mammalian cell transcription activator fusion protein, tTa (tetR-VP16), with the tetO-bearing minimal promoter derived from the humancytomegalovirus (hCMV) major immediate-early promoter to create atetR-tet operator system to control gene expression in mammalian cells.In one embodiment, a tetracycline inducible switch is used (Yao et al.,Human Gene Therapy; Gossen et al., Natl. Acad. Sci. USA, 89:5547-5551(1992); Shockett et al., Proc. Natl. Acad. Sci. USA, 92:6522-6526(1995)).

Additionally, the vector can contain, for example, some or all of thefollowing: a selectable marker gene, such as the neomycin gene forselection of stable or transient transfectants in mammalian cells;enhancer/promoter sequences from the immediate early gene of human CMVfor high levels of transcription; transcription termination and RNAprocessing signals from SV40 for mRNA stability; SV40 polyoma origins ofreplication and ColE1 for proper episomal replication; internal ribosomebinding sites (IRESes), versatile multiple cloning sites; and T7 and SP6RNA promoters for in vitro transcription of sense and antisense RNA.Suitable vectors and methods for producing vectors containing transgenesare well known and available in the art.

An expression vector comprising the nucleic acid can be transferred to ahost cell by conventional techniques (e.g., electroporation, liposomaltransfection, and calcium phosphate precipitation) and the transfectedcells are then cultured by conventional techniques to produce thecompounds described herein. In some embodiments, the expression of thecompounds described herein is regulated by a constitutive, an inducibleor a tissue-specific promoter.

The host cells used to express the compounds described herein orpolypeptides described herein may be either bacterial cells such asEscherichia coli, or, preferably, eukaryotic cells. In particular,mammalian cells, such as Chinese hamster ovary cells (CHO), inconjunction with a vector such as the major intermediate early genepromoter element from human cytomegalovirus is an effective expressionsystem for immunoglobulins (Foccking et al. (1986) “Powerful AndVersatile Enhancer-Promoter Unit For Mammalian Expression Vectors,” Gene45:101-106; Cockett et al. (1990) “High Level Expression Of TissueInhibitor Of Metalloproteinases In Chinese Hamster Ovary Cells UsingGlutamine Synthetase Gene Amplification,” Biotechnology 8:662-667).

A variety of host-expression vector systems may be utilized to expressthe compounds described herein or polypeptides described herein. Suchhost-expression systems represent vehicles by which the coding sequencesof the compounds described herein or polypeptides described herein maybe produced and subsequently purified, but also represent cells whichmay, when transformed or transfected with the appropriate nucleotidecoding sequences, express the compounds described herein in situ. Theseinclude, but are not limited to, microorganisms such as bacteria (e.g.,E. coli and B. subtilis) transformed with recombinant bacteriophage DNA,plasmid DNA or cosmid DNA expression vectors containing coding sequencesfor the compounds described herein; yeast (e.g., Saccharomyces pichia)transformed with recombinant yeast expression vectors containingsequences encoding the compounds described herein; insect cell systemsinfected with recombinant virus expression vectors (e.g., baclovirus)containing the sequences encoding the compounds described herein; plantcell systems infected with recombinant virus expression vectors (e.g.,cauliflower mosaic virus (CaMV) and tobacco mosaic virus (TMV) ortransformed with recombinant plasmid expression vectors (e.g., Tiplasmid) containing sequences encoding the molecules compounds describedherein; or mammalian cell systems (e.g., COS, CHO, BHK, 293, 293T, 3T3cells, lymphotic cells (see U.S. Pat. No. 5,807,715), Per C.6 cells(human retinal cells developed by Crucell) harboring recombinantexpression constructs containing promoters derived from the genome ofmammalian cells (e.g., metallothionein promoter) or from mammalianviruses (e.g., the adenovirus late promoter; the vaccinia virus 7.5Kpromoter).

In bacterial systems, a number of expression vectors may beadvantageously selected depending upon the use intended for the compoundbeing expressed. For example, when a large quantity of such a protein isto be produced, for the generation of pharmaceutical compositions ofcompounds described herein, vectors which direct the expression of highlevels of fusion protein products that are readily purified may bedesirable. Such vectors include, but arc not limited, to the E. coliexpression vector pUR278 (Rüther et al. (1983) “Easy Identification OfcDNA Clones,” EMBO J. 2:1791-1794), in which the coding sequence may beligated individually into the vector in frame with the lac Z codingregion so that a fusion protein is produced; pIN vectors (Inouye et al.(1985) “Up-Promoter Mutations In The 1pp Gene Of Escherichia Coli,”Nucleic Acids Res. 13:3101-3110; Van Heeke et al. (1989) “Expression OfHuman Asparagine Synthetase In Escherichia Coli,” J. Biol. Chem.24:5503-5509); and the like. pGEX vectors may also be used to expressforeign polypeptides as fusion proteins with glutathione S-transferase(GST). In general, such fusion proteins are soluble and can easily bepurified from lysed cells by adsorption and binding to a matrixglutathione-agarose beads followed by elution in the presence of freeglutathione. The pGEX vectors are designed to include thrombin or factorXa protease cleavage sites so that the cloned target gene product can bereleased from the GST moiety.

In an insect system, Autographa californica nuclear polyhedrosis virus(AcNPV) is used as a vector to express foreign genes. The virus grows inSpodoptera frugiperda cells. The coding sequence may be clonedindividually into non-essential regions (e.g., the polyhedrin gene) ofthe virus and placed under control of an AcNPV promoter (e.g., thepolyhedrin promoter).

In mammalian host cells, a number of viral-based expression systems maybe utilized. In cases where an adenovirus is used as an expressionvector, the coding sequence of interest may be ligated to an adenovirustranscription/translation control complex, e.g., the late promoter andtripartite leader sequence. This chimeric gene may then be inserted inthe adenovirus genome by in vitro or in vivo recombination. Insertion ina non-essential region of the viral genome (e.g., region E1 or E3) willresult in a recombinant virus that is viable and capable of expressingthe immunoglobulin molecule in infected hosts (e.g., see Logan et al.(1984) “Adenovirus Tripartite Leader Sequence Enhances Translation OfmRNAs Late After Infection,” Proc. Natl. Acad. Sci. USA 81:3655-3659).Specific initiation signals may also be required for efficienttranslation of inserted antibody coding sequences. These signals includethe ATG initiation codon and adjacent sequences. Furthermore, theinitiation codon must be in phase with the reading frame of the desiredcoding sequence to ensure translation of the entire insert. Theseexogenous translational control signals and initiation codons can be ofa variety of origins, both natural and synthetic. The efficiency ofexpression may be enhanced by the inclusion of appropriate transcriptionenhancer elements, transcription terminators, etc. (see Bitter et al.(1987) “Expression And Secretion Vectors For Yeast,” Methods in Enzymol.153:516-544).

In addition, a host cell strain may be chosen which modulates theexpression of the inserted sequences, or modifies and processes the geneproduct in the specific fashion desired. Such modifications (e.g.,glycosylation) and processing (e.g., cleavage) of protein products maybe important for the function of the protein. For example, in certainembodiments, the compounds described herein may be expressed as a singlegene product (e.g., as a single polypeptide chain, i.e., as apolyprotein precursor), requiring proteolytic cleavage by native orrecombinant cellular mechanisms to form separate polypeptides of thecompounds described herein. The disclosure thus encompasses engineeringa nucleic acid sequence to encode a polyprotein precursor moleculecomprising the polypeptides of the compounds described herein, whichincludes coding sequences capable of directing post translationalcleavage of said polyprotein precursor. Post-translational cleavage ofthe polyprotein precursor results in the polypeptides of the compoundsdescribed herein. The post translational cleavage of the precursormolecule comprising the polypeptides of the compounds described hereinmay occur in vivo (i.e., within the host cell by native or recombinantcell systems/mechanisms, e.g. furin cleavage at an appropriate site) ormay occur in vitro (e.g. incubation of said polypeptide chain in acomposition comprising proteases or peptidases of known activity and/orin a composition comprising conditions or reagents known to foster thedesired proteolytic action). Purification and modification ofrecombinant proteins is well known in the art such that the design ofthe polyprotein precursor could include a number of embodiments readilyappreciated by a skilled worker. Any known proteases or peptidases knownin the art can be used for the described modification of the precursormolecule, e.g., thrombin or factor Xa (Nagai et al. (1985) “OxygenBinding Properties Of Human Mutant Hemoglobins Synthesized InEscherichia Coli,” Proc. Nat. Acad. Sci. USA 82:7252-7255, and reviewedin Jenny et al. (2003) “A Critical Review Of The Methods For Cleavage OfFusion Proteins With Thrombin And Factor Xa,” Protein Expr. Purif.31:1-11, each of which is incorporated by reference herein in itsentirety)), enterokinase (Collins-Racie et al. (1995) “Production OfRecombinant Bovine Enterokinase Catalytic Subunit In Escherichia ColiUsing The Novel Secretory Fusion Partner DsbA,” Biotechnology 13:982-987hereby incorporated by reference herein in its entirety)), furin, andAcTEV (Parks et al. (1994) “Release Of Proteins And Peptides From FusionProteins Using A Recombinant Plant Virus Proteinase,” Anal. Biochem.216:413-417 hereby incorporated by reference herein in its entirety))and the Foot and Mouth Disease Virus Protease C3.

Different host cells have characteristic and specific mechanisms for thepost-translational processing and modification of proteins and geneproducts. Appropriate cell lines or host systems can be chosen to ensurethe correct modification and processing of the foreign proteinexpressed. To this end, eukaryotic host cells which possess the cellularmachinery for proper processing of the primary transcript,glycosylation, and phosphorylation of the gene product may be used. Suchmammalian host cells include but are not limited to CHO, VERY, BHK,HeLa, COS, MDCK, 293, 293T, 3T3, W138, BT483, Hs578T, HTB2, BT20 andT47D, CRL7030 and Hs578Bst.

For long-term, high-yield production of recombinant proteins, stableexpression is preferred. For example, cell lines which stably expresscompounds described herein may be engineered. Rather than usingexpression vectors which contain viral origins of replication, hostcells can be transformed with DNA controlled by appropriate expressioncontrol elements (e.g., promoter, enhancer, sequences, transcriptionterminators, polyadenylation sites, etc.), and a selectable marker.Following the introduction of the foreign DNA, engineered cells may beallowed to grow for 1-2 days in an enriched media, and then are switchedto a selective media. The selectable marker in the recombinant plasmidconfers resistance to the selection and allows cells to stably integratethe plasmid into their chromosomes and grow to form foci which in turncan be cloned and expanded into cell lines. This method mayadvantageously be used to engineer cell lines which express thecompounds described herein. Such engineered cell lines may beparticularly useful in screening and evaluation of compounds thatinteract directly or indirectly with the compounds described herein.

A number of selection systems may be used, including but not limited tothe herpes simplex virus thymidine kinase (Wigler et al. (1977)“Transfer Of Purified Herpes Virus Thymidine Kinase Gene To CulturedMouse Cells,” Cell 11: 223-232), hypoxanthine-guaninephosphoribosyltransferase (Szybalska et al. (1992) “Use Of The HPRT GeneAnd The HAT Selection Technique In DNA-Mediated Transformation OfMammalian Cells First Steps Toward Developing Hybridoma Techniques AndGene Therapy,” Bioessays 14: 495-500), and adeninephosphoribosyltransferase (Lowy et al. (1980) “Isolation Of TransformingDNA: Cloning The Hamster aprt Gene,” Cell 22: 817-823) genes can beemployed in tk-, hgprt- or aprt-cells, respectively. Also,antimetabolite resistance can be used as the basis of selection for thefollowing genes: dhfr, which confers resistance to methotrexate (Wigleret al. (1980) “Transformation Of Mammalian Cells With An AmplifiableDominant-Acting Gene,” Proc. Natl. Acad. Sci. USA 77:3567-3570; O'Hareet al. (1981) “Transformation Of Mouse Fibroblasts To MethotrexateResistance By A Recombinant Plasmid Expressing A ProkaryoticDihydrofolate Reductase,” Proc. Natl. Acad. Sci. USA 78: 1527-1531);gpt, which confers resistance to mycophenolic acid (Mulligan et al.(1981) “Selection For Animal Cells That Express The Escherichia coliGene Coding For Xanthine-Guanine Phosphoribosyltransferase,” Proc. Natl.Acad. Sci. USA 78: 2072-2076); neo, which confers resistance to theaminoglycoside G-418 (Tolstoshev (1993) “Gene Therapy, Concepts, CurrentTrials And Future Directions,” Ann. Rev. Pharmacol. Toxicol. 32:573-596;Mulligan (1993) “The Basic Science Of Gene Therapy,” Science260:926-932; and Morgan et al. (1993) “Human Gene Therapy,” Ann. Rev.Biochem. 62:191-217) and hygro, which confers resistance to hygromycin(Santerre et al. (1984) “Expression Of Prokaryotic Genes For HygromycinB And G418 Resistance As Dominant-Selection Markers In Mouse L Cells,”Gene 30:147-156). Methods commonly known in the art of recombinant DNAtechnology which can be used are described in Ausubel et al. (eds.),1993, Current Protocols in Molecular Biology, John Wiley & Sons, NY;Kriegler, 1990, Gene Transfer and Expression, A Laboratory Manual,Stockton Press, NY; and in Chapters 12 and 13, Dracopoli et al. (eds),1994, Current Protocols in Human Genetics, John Wiley & Sons, NY.;Colberre-Garapin et al. (1981) “A New Dominant Hybrid Selective MarkerFor Higher Eukaryotic Cells,” J. Mol. Biol. 150:1-14.

The expression levels of compounds described herein or polypeptidesdescribed herein can be increased by vector amplification (for a review,see Bebbington and Hentschel, The use of vectors based on geneamplification for the expression of cloned genes in mammalian cells inDNA cloning, Vol. 3 (Academic Press, New York, 1987). When a marker inthe vector system expressing a compound described herein is amplifiable,increase in the level of inhibitor present in culture of host cell willincrease the number of copies of the marker gene. Since the amplifiedregion is associated with the nucleotide sequence of a compounddescribed herein or a polypeptide described herein, production of thepolypeptide will also increase (Crouse et al. (1983) “Expression AndAmplification Of Engineered Mouse Dihydrofolate Reductase Minigenes,”Mol. Cell. Biol. 3:257-266).

The host cell may be co-transfected with two expression vectors, thefirst vector encoding the first polypeptide of a compound describedherein and the second vector encoding the second polypeptide of acompound described herein. The two vectors may contain identicalselectable markers which enable equal expression of both polypeptides.Alternatively, a single vector may be used which encodes bothpolypeptides. The coding sequences for the polypeptides of compoundsdescribed herein may comprise cDNA or genomic DNA.

Once a compound described herein or polypeptide described herein hasbeen recombinantly expressed, it may be purified by any method known inthe art for purification of polypeptides, polyproteins or antibodies(e.g., analogous to antibody purification schemes based on antigenselectivity) for example, by chromatography (e.g., ion exchange,affinity, particularly by affinity for the specific antigen (optionallyafter Protein A selection where the compound comprises an Fc domain (orportion thereof)), and sizing column chromatography), centrifugation,differential solubility, or by any other standard technique for thepurification of polypeptides or antibodies.

Other aspects of the disclosure relate to a cell comprising a nucleicacid described herein or a vector described herein. The cell may be aprokaryotic or eukaryotic cell. In some embodiments, the cell in amammalian cell. Exemplary cell types are described herein.

Yet other aspects of the disclosure relate to a method of producing acompound described herein or a polypeptide described herein (e.g., afirst polypeptide or a second polypeptide), the method comprisingobtaining a cell described herein and expressing nucleic acid describedherein in said cell. In some embodiments, the method further comprisesisolating and purifying a compound described herein or a polypeptidedescribed herein.

Methods of Treatment and Compositions for Use in Medicine

Other aspects of the disclosure relate to methods of treatment andcompositions for use in medicine. Non-limiting examples of compounds foruse in such methods and composition are those that comprise:

-   -   (i) a first polypeptide comprises the amino acid sequence of SEQ        ID NO:13 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:14;    -   (ii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:15 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:16;    -   (iii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:17 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:18;    -   (iv) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:19 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:20;    -   (v) a first polypeptide comprises the amino acid sequence of SEQ        ID NO:21 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:22;    -   (vi) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:23 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:24;    -   (vii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:25 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:26;    -   (viii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:27 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:28;    -   (ix) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:29 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:30;    -   (x) a first polypeptide comprises the amino acid sequence of SEQ        ID NO:31 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:32;    -   (xi) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:33 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:34; or    -   (xii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:35 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:36.

In some embodiments, the method of treatment or the use is a method oftreating an autoimmune or an inflammatory disease or use in such amethod. In some embodiments, the method comprises administering acompound described herein or a pharmaceutical composition comprisingsaid compound to a subject, e.g., a subject having or at risk for havingan autoimmune or an inflammatory disease.

The subject to be treated by the methods described herein can be amammal, more preferably a human. Mammals include, but are not limitedto, farm animals, sport animals, pets, primates, horses, dogs, cats,mice and rats. A human subject who needs the treatment may be a humansubject having, at risk for, or suspected of having a disease. A subjecthaving a disease can be identified by routine medical examination, e.g.,a physical examination, a laboratory test, an organ functional test, aCT scan, or an ultrasound. A subject suspected of having any of such adisease might show one or more symptoms of the disease. Signs andsymptoms for diseases, e.g., autoimmune and inflammatory diseases, arewell known to those of ordinary skill in the art. A subject at risk forthe disease can be a subject having one or more of the risk factors forthat disease.

Non-limiting examples of autoimmune diseases include rheumatoidarthritis, psoriasis, type 1 diabetes, systemic lupus erythematosus,transplant rejection, autoimmune thyroid disease (Hashimoto's disease),sarcoidosis, scleroderma, granulomatous vasculitis, Crohn's disease,ulcerative colitis, Sjogren's disease, ankylosing spondylitis, psoriaticarthritis, polymyositis dermatomyositis, polyarteritis nodosa,immunologically mediated blistering skin diseases, Behcet's syndrome,multiple sclerosis, systemic sclerosis, Goodpasture's disease or immunemediated glomerulonephritis.

Non-limiting examples of inflammatory diseases include includingrheumatoid arthritis, systemic lupus erythematosus, alopecia areata,anklosing spondylitis, antiphospholipid syndrome, autoimmune Addison'sdisease, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmuneinner ear disease, autoimmune lymphoproliferative syndrome (ALPS),autoimmune thrombocytopenic purpura (ATP), Behcet's disease, bullouspemphigoid, cardiomyopathy, celiac sprue-dermatitis, chronic fatiguesyndrome immune deficiency syndrome (CFIDS), chronic inflammatorydemyelinating polyneuropathy, cicatricial pemphigoid, cold agglutinindisease, Crest syndrome, Crohn's disease, Dego's disease,dermatomyasitis, dermatomyositis-juvenile, discoid lupus, essentialmixed cryoglobulinemia, fibromyalgia-fibromyositis, grave's disease,Guillain-Barre, hashimoto's thyroiditis, idiopathic pulmonary fibrosis,idiopathic thrombocytopenia purpura (ITP), Iga nephropathy, insulindependent diabetes (Type I), juvenile arthritis, Meniere's disease,mixed connective tissue disease, multiple sclerosis, myasthenia gravis,pemphigus vulgaris, pernicious anemia, polyarteritis nodosa,polychondritis, polyglancular syndromes, polymyalgia rheumatica,polymyositis and dermatomyositis, primary agammaglobulinemia, primarybiliary cirrhosis, psoriasis, Raynaud's phenomenon, Reiter's syndrome,rheumatic fever, sarcoidosis, scleroderma, Sjogren's syndrome, stiff-mansyndrome, Takayasu arteritis, temporal arteritis/giant cell arteritis,ulcerative colitis, uveitis, vasculitis, vitiligo, and Wegener'sgranulomatosis. In some embodiments, the autoimmune or inflammatorydisease is Crohn's disease, ankylosing spondylitis, or psoriaticarthritis.

To practice a method disclosed herein, an effective amount of a compoundor pharmaceutical composition described herein can be administered to asubject (e.g., a human) in need of the treatment. Various deliverysystems are known and can be used to administer the compounds of theinvention. Methods of administration include, but are not limited to,intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous,intranasal, epidural, and oral routes. The compounds of the inventioncan be administered, for example by infusion, bolus or injection, andcan be administered together with other biologically active agents suchas anti-inflammatory agents. Administration can be systemic or local. Inpreferred embodiments, the administration is by subcutaneous injection.Formulations for such injections may be prepared in, for example,prefilled syringes that may be administered once every other week.

“An effective amount” as used herein refers to the amount of eachcompound required to confer therapeutic effect on the subject, eitheralone or in combination with one or more other compounds. Effectiveamounts vary, as recognized by those skilled in the art, depending onthe particular condition being treated, the severity of the condition,the individual subject parameters including age, physical condition,size, gender and weight, the duration of the treatment, the nature ofconcurrent therapy (if any), the specific route of administration andlike factors within the knowledge and expertise of the healthpractitioner. These factors are well known to those of ordinary skill inthe art and can be addressed with no more than routine experimentation.It is generally preferred that a maximum dose of the individualcomponents or combinations thereof be used, that is, the highest safedose according to sound medical judgment. It will be understood by thoseof ordinary skill in the art, however, that a subject may insist upon alower dose or tolerable dose for medical reasons, psychological reasonsor for virtually any other reasons.

Empirical considerations, such as the half-life, generally willcontribute to the determination of the dosage. For example, compoundsthat are compatible with the human immune system, such as compoundscomprising regions from humanized antibodies or fully human antibodies,may be used to prolong half-life of the compound and to prevent thecompound being attacked by the host's immune system. Frequency ofadministration may be determined and adjusted over the course oftherapy, and is generally, but not necessarily, based on treatmentand/or suppression and/or amelioration and/or delay of a disease.Alternatively, sustained continuous release formulations of a compoundmay be appropriate. Various formulations and devices for achievingsustained release are known in the art.

In some embodiments, dosage is daily, every other day, every three days,every four days, every five days, or every six days. In someembodiments, dosing frequency is once every week, every 2 weeks, every 4weeks, every 5 weeks, every 6 weeks, every 7 weeks, every 8 weeks, every9 weeks, or every 10 weeks; or once every month, every 2 months, orevery 3 months, or longer. The progress of this therapy is easilymonitored by conventional techniques and assays. The dosing regimen(including the compound used) can vary over time.

In some embodiments, for an adult subject of normal weight, dosesranging from about 0.01 to 1000 mg/kg may be administered. In someembodiments, the dose is between 1 to 200 mg. The particular dosageregimen, i.e., dose, timing and repetition, will depend on theparticular subject and that subject's medical history, as well as theproperties of the compound (such as the half-life of the compound, andother considerations well known in the art).

For the purpose of the present disclosure, the appropriate dosage of acompound as described herein will depend on the specific compound (orcompositions thereof) employed, the formulation and route ofadministration, the type and severity of the disease, whether thecompound is administered for preventive or therapeutic purposes,previous therapy, the subject's clinical history and response to theantagonist, and the discretion of the attending physician. Typically theclinician will administer a compound until a dosage is reached thatachieves the desired result. Administration of one or more compounds canbe continuous or intermittent, depending, for example, upon therecipient's physiological condition, whether the purpose of theadministration is therapeutic or prophylactic, and other factors knownto skilled practitioners. The administration of a compound may beessentially continuous over a preselected period of time or may be in aseries of spaced dose, e.g., either before, during, or after developinga disease.

As used herein, the term “treating” refers to the application oradministration of a compound or composition including the compound to asubject, who has a disease, a symptom of the disease, or apredisposition toward the disease, with the purpose to cure, heal,alleviate, relieve, alter, remedy, ameliorate, improve, or affect thedisease, the symptom of the disease, or the predisposition toward thedisease.

Alleviating a disease includes delaying the development or progressionof the disease, or reducing disease severity. Alleviating the diseasedoes not necessarily require curative results. As used therein,“delaying” the development of a disease means to defer, hinder, slow,retard, stabilize, and/or postpone progression of the disease. Thisdelay can be of varying lengths of time, depending on the history of thedisease and/or individuals being treated. A method that “delays” oralleviates the development of a disease, or delays the onset of thedisease, is a method that reduces probability of developing one or moresymptoms of the disease in a given time frame and/or reduces extent ofthe symptoms in a given time frame, when compared to not using themethod. Such comparisons are typically based on clinical studies, usinga number of subjects sufficient to give a statistically significantresult.

“Development” or “progression” of a disease means initial manifestationsand/or ensuing progression of the disease. Development of the diseasecan be detectable and assessed using standard clinical techniques aswell known in the art. However, development also refers to progressionthat may be undetectable. For purpose of this disclosure, development orprogression refers to the biological course of the symptoms.“Development” includes occurrence, recurrence, and onset. As used herein“onset” or “occurrence” of a disease includes initial onset and/orrecurrence.

In some embodiments, the compound described herein is administered to asubject in need of the treatment at an amount sufficient to inhibit theactivity of one or both of TNF-alpha or IL23A by at least 20% (e.g.,30%, 40%, 50%, 60%, 70%, 80%, 90% or greater) in vivo or in vitro.Methods for determining the inhibitory capability of a compound areknown in the art. Exemplary TNF-alpha and IL23A inhibition assays areprovided in the Examples.

Conventional methods, known to those of ordinary skill in the art ofmedicine, can be used to administer the compound or pharmaceuticalcomposition to the subject, depending upon the type of disease to betreated or the site of the disease. This composition can also beadministered via other conventional routes, e.g., administered orally,parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir. The term “parenteral”as used herein includes subcutaneous, intracutaneous, intravenous,intramuscular, intraarticular, intraarterial, intrasynovial,intrasternal, intrathecal, intralesional, and intracranial injection orinfusion techniques. In addition, it can be administered to the subjectvia injectable depot routes of administration such as using 1-, 3-, or6-month depot injectable or biodegradable materials and methods.

Pharmaceutical Compositions

Yet other aspects of the disclosure relate to pharmaceuticalcompositions comprising a compound described herein. A compositioncomprising a compound of the invention (e.g., compounds specific forboth TNF-alpha and IL23A) can be administered to a subject having or atrisk of having an autoimmune or an inflammatory disease. The inventionfurther provides for the use of a compound of the invention in themanufacture of a medicament for treatment of an autoimmune or aninflammatory disease. The compounds can be administered either alone orin combination with other compositions in the prevention or treatment ofan autoimmune or an inflammatory disease. Non-limiting examples ofcompounds of the invention for use in such pharmaceutical compositionsare those that comprise:

-   -   (i) a first polypeptide comprises the amino acid sequence of SEQ        ID NO:13 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:14;    -   (ii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:15 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:16;    -   (iii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:17 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:18;    -   (iv) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:19 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:20;    -   (v) a first polypeptide comprises the amino acid sequence of SEQ        ID NO:21 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:22;    -   (vi) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:23 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:24;    -   (vii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:25 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:26;    -   (viii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:27 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:28;    -   (ix) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:29 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:30;    -   (x) a first polypeptide comprises the amino acid sequence of SEQ        ID NO:31 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:32;    -   (xi) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:33 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:34; or    -   (xii) a first polypeptide comprises the amino acid sequence of        SEQ ID NO:35 and a second polypeptide comprises the amino acid        sequence of SEQ ID NO:36.

As used herein, the term “pharmaceutical composition” refers to theformulation of a compound described herein in combination with apharmaceutically acceptable carrier. The pharmaceutical composition canfurther comprise additional agents (e.g. for specific delivery,increasing half-life, or other therapeutic compounds).

As used here, the term “pharmaceutically-acceptable carrier” means apharmaceutically-acceptable material, composition or vehicle, such as aliquid or solid filler, diluent, excipient, manufacturing aid (e.g.,lubricant, talc magnesium, calcium or zinc stearate, or steric acid), orsolvent encapsulating material, involved in carrying or transporting thecompound from one site (e.g., the delivery site) of the body, to anothersite (e.g., organ, tissue or portion of the body). A pharmaceuticallyacceptable carrier is “acceptable” in the sense of being compatible withthe other ingredients of the formulation and not injurious to the tissueof the subject (e.g., physiologically compatible, sterile, physiologicpH, etc.). Some examples of materials which can serve aspharmaceutically-acceptable carriers include: (1) sugars, such aslactose, glucose and sucrose; (2) starches, such as corn starch andpotato starch; (3) cellulose, and its derivatives, such as sodiumcarboxymethyl cellulose, methylcellulose, ethyl cellulose,microcrystalline cellulose and cellulose acetate; (4) powderedtragacanth; (5) malt; (6) gelatin; (7) lubricating agents, such asmagnesium stearate, sodium lauryl sulfate and talc; (8) excipients, suchas cocoa butter and suppository waxes; (9) oils, such as peanut oil,cottonseed oil, safflower oil, sesame oil, olive oil, corn oil andsoybean oil; (10) glycols, such as propylene glycol; (11) polyols, suchas glycerin, sorbitol, mannitol and polyethylene glycol (PEG); (12)esters, such as ethyl oleate and ethyl laurate; (13) agar; (14)buffering agents, such as magnesium hydroxide and aluminum hydroxide;(15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18)Ringer's solution; (19) ethyl alcohol; (20) pH buffered solutions; (21)polyesters, polycarbonates and/or polyanhydrides; (22) bulking agents,such as polypeptides and amino acids (23) serum component, such as serumalbumin, HDL and LDL; (22) C2-C12 alcohols, such as ethanol; and (23)other non-toxic compatible substances employed in pharmaceuticalformulations. Wetting agents, coloring agents, release agents, coatingagents, sweetening agents, flavoring agents, perfuming agents,preservative and antioxidants can also be present in the formulation.The terms such as “excipient”, “carrier”, “pharmaceutically acceptablecarrier” or the like are used interchangeably herein.

In some embodiments, a compound of the invention in a composition isadministered by injection, by means of a catheter, by means of asuppository, or by means of an implant, the implant being of a porous,non-porous, or gelatinous material, including a membrane, such as asialastic membrane, or a fiber. Typically, when administering thecomposition, materials to which the compound of the invention does notabsorb are used.

In other embodiments, the compounds of the invention are delivered in acontrolled release system. In one embodiment, a pump may be used (see,e.g., Langer, 1990, Science 249:1527-1533; Sefton, 1989, CRC Crit. Ref.Biomed. Eng. 14:201; Buchwald et al., 1980, Surgery 88:507; Saudek etal., 1989, N. Engl. J. Med. 321:574). In another embodiment, polymericmaterials can be used. (See, e.g., Medical Applications of ControlledRelease (Langer and Wise eds., CRC Press, Boca Raton, Fla., 1974);Controlled Drug Bioavailability, Drug Product Design and Performance(Smolen and Ball eds., Wiley, New York, 1984); Ranger and Peppas, 1983,Macromol. Sci. Rev. Macromol. Chem. 23:61. See also Levy et al., 1985,Science 228:190; During et al., 1989, Ann. Neurol. 25:351; Howard etal., 1989, J. Neurosurg. 71:105.) Other controlled release systems arediscussed, for example, in Langer, supra.

Compounds of the invention can be administered as pharmaceuticalcompositions comprising a therapeutically effective amount of a bindingagent and one or more pharmaceutically compatible ingredients.

In typical embodiments, the pharmaceutical composition is formulated inaccordance with routine procedures as a pharmaceutical compositionadapted for intravenous or subcutaneous administration to a subject,e.g., a human being. Typically, compositions for administration byinjection are solutions in sterile isotonic aqueous buffer. Wherenecessary, the pharmaceutical can also include a solubilizing agent anda local anesthetic such as lignocaine to ease pain at the site of theinjection. Generally, the ingredients are supplied either separately ormixed together in unit dosage form, for example, as a dry lyophilizedpowder or water free concentrate in a hermetically sealed container suchas an ampoule or sachette indicating the quantity of active agent. Wherethe pharmaceutical is to be administered by infusion, it can bedispensed with an infusion bottle containing sterile pharmaceuticalgrade water or saline. Where the pharmaceutical is administered byinjection, an ampoule of sterile water for injection or saline can beprovided so that the ingredients can be mixed prior to administration.

A pharmaceutical composition for systemic administration may be aliquid, e.g., sterile saline, lactated Ringer's or Hank's solution. Inaddition, the pharmaceutical composition can be in solid forms andre-dissolved or suspended immediately prior to use. Lyophilized formsare also contemplated.

The pharmaceutical composition can be contained within a lipid particleor vesicle, such as a liposome or microcrystal, which is also suitablefor parenteral administration. The particles can be of any suitablestructure, such as unilamellar or plurilamellar, so long as compositionsare contained therein. Compounds can be entrapped in ‘stabilizedplasmid-lipid particles’ (SPLP) containing the fusogenic lipiddioleoylphosphatidylethanolamine (DOPE), low levels (5-10 mol %) ofcationic lipid, and stabilized by a polyethyleneglycol (PEG) coating(Zhang Y. P. et al., Gene Ther. 1999, 6:1438-47). Positively chargedlipids such asN-[1-(2,3-dioleoyloxi)propyl]-N,N,N-trimethyl-amoniummethylsulfate, or“DOTAP,” are particularly preferred for such particles and vesicles. Thepreparation of such lipid particles is well known. See, e.g., U.S. Pat.Nos. 4,880,635; 4,906,477; 4,911,928; 4,917,951; 4,920,016; and4,921,757.

The pharmaceutical compositions of this disclosure may be administeredor packaged as a unit dose, for example. The term “unit dose” when usedin reference to a pharmaceutical composition of the present disclosurerefers to physically discrete units suitable as unitary dosage for thesubject, each unit containing a predetermined quantity of activematerial calculated to produce the desired therapeutic effect inassociation with the required diluent; i.e., carrier, or vehicle.

In some embodiments, a compound described herein may be conjugated to atherapeutic moiety, e.g., an anti-inflammatory agent. Techniques forconjugating such therapeutic moieties to polypeptides, including e.g.,Fc domains, are well known; see, e.g., Amon et al., “MonoclonalAntibodies For Immunotargeting Of Drugs In Cancer Therapy”, inMonoclonal Antibodies And Cancer Therapy, Reisfeld et al. (eds.), 1985,pp. 243-56, Alan R. Liss, Inc.); Hellstrom et al., “Antibodies For DrugDelivery”, in Controlled Drug Delivery (2nd Ed.), Robinson et al.(eds.), 1987, pp. 623-53, Marcel Dekker, Inc.); Thorpe, “AntibodyCarriers Of Cytotoxic Agents In Cancer Therapy: A Review”, in MonoclonalAntibodies '84: Biological And Clinical Applications, Pinchera et al.(eds.), 1985, pp. 475-506); “Analysis, Results, And Future ProspectiveOf The Therapeutic Use Of Radiolabeled Antibody In Cancer Therapy”, inMonoclonal Antibodies For Cancer Detection And Therapy, Baldwin et al.(eds.), 1985, pp. 303-16, Academic Press; and Thorpe et al. (1982) “ThePreparation And Cytotoxic Properties Of Antibody-Toxin Conjugates,”Immunol. Rev., 62:119-158.

Further, the pharmaceutical composition can be provided as apharmaceutical kit comprising (a) a container containing a compound ofthe invention in lyophilized form and (b) a second container containinga pharmaceutically acceptable diluent (e.g., sterile water) forinjection. The pharmaceutically acceptable diluent can be used forreconstitution or dilution of the lyophilized compound of the invention.Optionally associated with such container(s) can be a notice in the formprescribed by a governmental agency regulating the manufacture, use orsale of pharmaceuticals or biological products, which notice reflectsapproval by the agency of manufacture, use or sale for humanadministration.

In another aspect, an article of manufacture containing materials usefulfor the treatment of the diseases described above is included. In someembodiments, the article of manufacture comprises a container and alabel. Suitable containers include, for example, bottles, vials,syringes, and test tubes. The containers may be formed from a variety ofmaterials such as glass or plastic. In some embodiments, the containerholds a composition that is effective for treating a disease describedherein and may have a sterile access port. For example, the containermay be an intravenous solution bag or a vial having a stopper pierceableby a hypodermic injection needle. The active agent in the composition isa compound of the invention. In some embodiments, the label on orassociated with the container indicates that the composition is used fortreating the disease of choice. The article of manufacture may furthercomprise a second container comprising a pharmaceutically-acceptablebuffer, such as phosphate-buffered saline, Ringer's solution, ordextrose solution. It may further include other materials desirable froma commercial and user standpoint, including other buffers, diluents,filters, needles, syringes, and package inserts with instructions foruse.

Without further elaboration, it is believed that one skilled in the artcan, based on the above description, utilize the present disclosure toits fullest extent. The following specific embodiments are, therefore,to be construed as merely illustrative, and not limitative of theremainder of the disclosure in any way whatsoever. All publicationscited herein are incorporated by reference for the purposes or subjectmatter referenced herein.

EXAMPLES Example 1. Construction of Exemplary Compounds Targeting IL23Aand TNF-Alpha

Table 2A below provides exemplary compounds that bind to both IL23A andTNF-alpha that were utilized in the Examples below. These compounds wereproduced by recombinant methods known in the art (see, e.g., PCTPublications WO 2006/113665, WO 2008/157379, and WO 2010/080538, all ofwhich are incorporated herein by reference). Briefly, plasmids encodingthe first and second polypeptide for each compound were transfectedtogether into CHO-S cells using FreeStyle MAX Reagent (CHO). The cellswere cultured for 13-14 days and the compounds produced by the cellswere purified using Protein-A chromatography. The compounds were furtherpurified using a size exclusion chromatography.

TABLE 2A Exemplary IL23A and TNF-alpha binding compounds Large LargeSmall Small Linker SEQ ID NO: Compound ID Chain vL Chain vH Chain vLChain vH types Isotype (1st/2nd) Compound A TNFa(1) IL23A(1) IL23A(1)TNFa(1) GS IgG1KO-YTE 13/14 VL (SEQ VH (SEQ VL (SEQ VH (SEQ ID NO: 2) IDNO: 7) ID NO: 8) ID NO: 1) Compound B TNFa(1) IL23A(1) IL23A(1) TNFa(1)VF IgG1KO-YTE 15/16 VL (SEQ VH (SEQ VL (SEQ VH (SEQ ID NO: 2) ID NO: 7)ID NO: 8) ID NO: 1) Compound C IL23A(1) TNFa(1) TNFa(1) IL23A(1) GSIgG1KO-YTE 17/18 VL (SEQ VH (SEQ VL (SEQ VH (SEQ ID NO: 8) ID NO: 1) IDNO: 2) ID NO: 7) Compound D IL23A(1) TNFa(1) TNFa(1) IL23A(1) VFIgG1KO-YTE 19/20 VL (SEQ VH (SEQ VL (SEQ VH (SEQ ID NO: 8) ID NO: 1) IDNO: 2) ID NO: 7) Compound E IL23A(1) TNFa(2) TNFa(2) IL23A(1) GSIgG1KO-YTE 21/22 VL (SEQ VH (SEQ VL (SEQ VH (SEQ ID NO: 8) ID NO: 3) IDNO: 4) ID NO: 7) Compound F IL23A(1) TNFa(2) TNFa(2) IL23A(1) VFIgG1KO-YTE 23/24 VL (SEQ VH (SEQ VL (SEQ VH (SEQ ID NO: 8) ID NO: 3) IDNO: 4) ID NO: 7) Compound G TNFa(2) IL23A(1) IL23A(1) TNFa(2) GSIgG1KO-YTE 25/26 VL (SEQ VH (SEQ VL (SEQ VH (SEQ ID NO: 4) ID NO: 7) IDNO: 8) ID NO: 3) Compound H TNFa(2) IL23A(1) IL23A(1) TNFa(2) VFIgG1KO-YTE 27/28 VL (SEQ VH (SEQ VL (SEQ VH (SEQ ID NO: 4) ID NO: 7) IDNO: 8) ID NO: 3) Compound I TNFa(3) IL23A(1) IL23A(1) TNFa(3) GSIgG1KO-YTE 29/30 VL (SEQ VH (SEQ VL (SEQ VH (SEQ ID NO: 6) ID NO: 7) IDNO: 8) ID NO: 5) Compound J IL23A(1) TNFa(3) TNFa(3) IL23A(1) GSIgG1KO-YTE 31/32 VL (SEQ VH (SEQ VL (SEQ VH (SEQ ID NO: 8) ID NO: 5) IDNO: 6) ID NO: 7) Compound K TNFa(3) IL23A(1) IL23A(1) TNFa(3) VFIgG1KO-YTE 33/34 VL (SEQ VH (SEQ VL (SEQ VH (SEQ ID NO: 6) ID NO: 7) IDNO: 8) ID NO: 5) Compound L IL23A(1) TNFa(3) TNFa(3) IL23A(1) VFIgG1KO-YTE 35/36 VL (SEQ VH (SEQ VL (SEQ VH (SEQ ID NO: 8) ID NO: 5) IDNO: 6) ID NO: 7) Compound M TNFa(1) IL23A(1) IL23A(1) TNFa(1) GS IgG1KO44/45 VL (SEQ VH (SEQ VL (SEQ VH (SEQ ID NO: 2) ID NO: 7) ID NO: 8) IDNO: 1) Compound N IL23A(1) TNFa(1) TNFa(1) IL23A(1) VF IgG1KO 46/47 VL(SEQ VH (SEQ VL (SEQ VH (SEQ ID NO: 8) ID NO: 1) ID NO: 2) ID NO: 7)Compound O IL23A(1) TNFa(2) TNFa(2) IL23A(1) GS IgG1KO 48/49 VL (SEQ VH(SEQ VL (SEQ VH (SEQ ID NO: 8) ID NO: 3) ID NO: 4) ID NO: 7) Compound PIL23A(1) TNFa(2) TNFa(2) IL23A(1) VF IgG1KO 50/51 VL (SEQ VH (SEQ VL(SEQ VH (SEQ ID NO: 8) ID NO: 3) ID NO: 4) ID NO: 7) Compound Q TNFa(2)IL23A(1) IL23A(1) TNFa(2) GS IgG4Pro 52/53 VL (SEQ VH (SEQ VL (SEQ VH(SEQ ID NO: 4) ID NO: 7) ID NO: 8) ID NO: 3) Compound R TNFa(2) IL23A(1)IL23A(1) TNFa(2) GS IgG4Pro-YTE 54/55 VL (SEQ VH (SEQ VL (SEQ VH (SEQ IDNO: 4) ID NO: 7) ID NO: 8) ID NO: 3) Compound S TNFa(2) IL23A(1)IL23A(1) TNFa(2) GS IgG4Pro 56/57 VL (SEQ VH (SEQ VL (SEQ VH (SEQ ID NO:4) ID NO: 7) ID NO: 8) ID NO: 3) Compound T TNFa(2) IL23A(1) IL23A(1)TNFa(2) GS IgG4Pro-YTE 58/59 VL (SEQ VH (SEQ VL (SEQ VH (SEQ ID NO: 4)ID NO: 7) ID NO: 8) ID NO: 3) Compound U TNFa(2) IL23A(1) IL23A(1)TNFa(2) VF IgG1KO 60/61 VL (SEQ VH (SEQ VL (SEQ VH (SEQ ID NO: 4) ID NO:7) ID NO: 8) ID NO: 3) Compound V TNFa(2) IL23A(1) IL23A(1) TNFa(2) VFIgG1WT 62/63 VL (SEQ VH (SEQ VL (SEQ VH (SEQ ID NO: 4) ID NO: 7) ID NO:8) ID NO: 3) Compound W TNFa(2) IL23A(1) IL23A(1) TNFa(2) VF IgG4Pro64/65 VL (SEQ VH (SEQ VL (SEQ VH (SEQ ID NO: 4) ID NO: 7) ID NO: 8) IDNO: 3) Compound X TNFa(2) IL23A(1) IL23A(1) TNFa(2) VF IgG4Pro-YTE 66/67VL (SEQ VH (SEQ VL (SEQ VH (SEQ ID NO: 4) ID NO: 7) ID NO: 8) ID NO: 3)Compound Y IL23A(1) TNFa(2) TNFa(2) IL23A(1) GS IgG1WT 68/69 VL (SEQ VH(SEQ VL (SEQ VH (SEQ ID NO: 8) ID NO: 3) ID NO: 4) ID NO: 7) Compound ZIL23A(1) TNFa(2) TNFa(2) IL23A(1) GS IgG4Pro 70/71 VL (SEQ VH (SEQ VL(SEQ VH (SEQ ID NO: 8) ID NO: 3) ID NO: 4) ID NO: 7) Compound AAIL23A(1) TNFa(2) TNFa(2) IL23A(1) GS IgG4Pro-YTE 72/73 VL (SEQ VH (SEQVL (SEQ VH (SEQ ID NO: 8) ID NO: 3) ID NO: 4) ID NO: 7) Compound ABIL23A(1) TNFa(2) TNFa(2) IL23A(1) VF IgG1WT 74/75 VL (SEQ VH (SEQ VL(SEQ VH (SEQ ID NO: 8) ID NO: 3) ID NO: 4) ID NO: 7) Compound ACIL23A(1) TNFa(2) TNFa(2) IL23A(1) VF IgG4Pro 76/77 VL (SEQ VH (SEQ VL(SEQ VH (SEQ ID NO: 8) ID NO: 3) ID NO: 4) ID NO: 7) Compound ADIL23A(1) TNFa(2) TNFa(2) IL23A(1) VF IgG4Pro-YTE 78/79 VL (SEQ VH (SEQVL (SEQ VH (SEQ ID NO: 8) ID NO: 3) ID NO: 4) ID NO: 7) Compound AETNFa(3) IL23A(1) IL23A(1) TNFa(3) GS IgG1KO 80/81 VL (SEQ VH (SEQ VL(SEQ VH (SEQ ID NO: 6) ID NO: 7) ID NO: 8) ID NO: 5) Compound AF TNFa(3)IL23A(1) IL23A(1) TNFa(3) GS IgG1WT 82/83 VL (SEQ VH (SEQ VL (SEQ VH(SEQ ID NO: 6) ID NO: 7) ID NO: 8) ID NO: 5) Compound AG TNFa(3)IL23A(1) IL23A(1) TNFa(3) GS IgG4Pro 84/85 VL (SEQ VH (SEQ VL (SEQ VH(SEQ ID NO: 6) ID NO: 7) ID NO: 8) ID NO: 5) Compound AH TNFa(3)IL23A(1) IL23A(1) TNFa(3) GS IgG4Pro-YTE 86/87 VL (SEQ VH (SEQ VL (SEQVH (SEQ ID NO: 6) ID NO: 7) ID NO: 8) ID NO: 5) Compound AI TNFa(3)IL23A(1) IL23A(1) TNFa(3) VF IgG1KO 88/89 VL (SEQ VH (SEQ VL (SEQ VH(SEQ ID NO: 6) ID NO: 7) ID NO: 8) ID NO: 5) Compound AJ TNFa(3)IL23A(1) IL23A(1) TNFa(3) VF IgG1WT 90/91 VL (SEQ VH (SEQ VL (SEQ VH(SEQ ID NO: 6) ID NO: 7) ID NO: 8) ID NO: 5) Compound AK TNFa(3)IL23A(1) IL23A(1) TNFa(3) VF IgG4Pro 92/93 VL (SEQ VH (SEQ VL (SEQ VH(SEQ ID NO: 6) ID NO: 7) ID NO: 8) ID NO: 5) Compound AL TNFa(3)IL23A(1) IL23A(1) TNFa(3) VF IgG4Pro-YTE 94/95 VL (SEQ VH (SEQ VL (SEQVH (SEQ ID NO: 6) ID NO: 7) ID NO: 8) ID NO: 5) Compound AM IL23A(1)TNFa(3) TNFa(3) IL23A(1) GS IgG1KO 96/97 VL (SEQ VH (SEQ VL (SEQ VH (SEQID NO: 8) ID NO: 5) ID NO: 6) ID NO: 7) Compound AN IL23A(1) TNFa(3)TNFa(3) IL23A(1) GS IgG1WT 98/99 VL (SEQ VH (SEQ VL (SEQ VH (SEQ ID NO:8) ID NO: 5) ID NO: 6) ID NO: 7) Compound AO IL23A(1) TNFa(3) TNFa(3)IL23A(1) GS IgG4Pro 100/101 VL (SEQ VH (SEQ VL (SEQ VH (SEQ ID NO: 8) IDNO: 5) ID NO: 6) ID NO: 7) Compound AP IL23A(1) TNFa(3) TNFa(3) IL23A(1)GS IgG4Pro-YTE 102/103 VL (SEQ VH (SEQ VL (SEQ VH (SEQ ID NO: 8) ID NO:5) ID NO: 6) ID NO: 7) Compound AQ IL23A(1) TNFa(3) TNFa(3) IL23A(1) VFIgG1KO 104/105 VL (SEQ VH (SEQ VL (SEQ VH (SEQ ID NO: 8) ID NO: 5) IDNO: 6) ID NO: 7) Compound AR IL23A(1) TNFa(3) TNFa(3) IL23A(1) VF IgG1WT106/107 VL (SEQ VH (SEQ VL (SEQ VH (SEQ ID NO: 8) ID NO: 5) ID NO: 6) IDNO: 7) Compound AS IL23A(1) TNFa(3) TNFa(3) IL23A(1) VF IgG4Pro 108/109VL (SEQ VH (SEQ VL (SEQ VH (SEQ ID NO: 8) ID NO: 5) ID NO: 6) ID NO: 7)Compound AT IL23A(1) TNFa(3) TNFa(3) IL23A(1) VF IgG4Pro-YTE 110/111 VL(SEQ VH (SEQ VL (SEQ VH (SEQ ID NO: 8) ID NO: 5) ID NO: 6) ID NO: 7)Compound AU TNFa(2) IL23A(1) IL23A(1) TNFa(2) GS IgG1KO 112/113 VL (SEQVH (SEQ VL (SEQ VH (SEQ ID NO: 4) ID NO: 7) ID NO: 8) ID NO: 3) CompoundAV TNFa(1) IL23A(1) IL23A(1) TNFa(1) GS IgG1WT 114/115 VL (SEQ VH (SEQVL (SEQ VH (SEQ ID NO: 2) ID NO: 7) ID NO: 8) ID NO: 1) Compound AWTNFa(1) IL23A(1) IL23A(1) TNFa(1) GS IgG4Pro 116/117 VL (SEQ VH (SEQ VL(SEQ VH (SEQ ID NO: 2) ID NO: 7) ID NO: 8) ID NO: 1) Compound AX TNFa(1)IL23A(1) IL23A(1) TNFa(1) GS IgG4Pro-YTE 118/119 VL (SEQ VH (SEQ VL (SEQVH (SEQ ID NO: 2) ID NO: 7) ID NO: 8) ID NO: 1) Compound AY TNFa(1)IL23A(1) IL23A(1) TNFa(1) VF IgG1KO 120/121 VL (SEQ VH (SEQ VL (SEQ VH(SEQ ID NO: 2) ID NO: 7) ID NO: 8) ID NO: 1) Compound AZ TNFa(1)IL23A(1) IL23A(1) TNFa(1) VF IgG1WT 122/123 VL (SEQ VH (SEQ VL (SEQ VH(SEQ ID NO: 2) ID NO: 7) ID NO: 8) ID NO: 1) Compound BA TNFa(1)IL23A(1) IL23A(1) TNFa(1) VF IgG4Pro 124/125 VL (SEQ VH (SEQ VL (SEQ VH(SEQ ID NO: 2) ID NO: 7) ID NO: 8) ID NO: 1) Compound BB TNFa(1)IL23A(1) IL23A(1) TNFa(1) VF IgG4Pro-YTE 126/127 VL (SEQ VH (SEQ VL (SEQVH (SEQ ID NO: 2) ID NO: 7) ID NO: 8) ID NO: 1) Compound BC IL23A(1)TNFa(1) TNFa(1) IL23A(1) GS IgG1KO 128/129 VL (SEQ VH (SEQ VL (SEQ VH(SEQ ID NO: 8) ID NO: 1) ID NO: 2) ID NO: 7) Compound BD IL23A(1)TNFa(1) TNFa(1) IL23A(1) GS IgG1WT 130/131 VL (SEQ VH (SEQ VL (SEQ VH(SEQ ID NO: 8) ID NO: 1) ID NO: 2) ID NO: 7) Compound BE IL23A(1)TNFa(1) TNFa(1) IL23A(1) GS IgG4Pro 132/133 VL (SEQ VH (SEQ VL (SEQ VH(SEQ ID NO: 8) ID NO: 1) ID NO: 2) ID NO: 7) Compound BF IL23A(1)TNFa(1) TNFa(1) IL23A(1) GS IgG4Pro-YTE 134/135 VL (SEQ VH (SEQ VL (SEQVH (SEQ ID NO: 8) ID NO: 1) ID NO: 2) ID NO: 7) Compound BG TNFa(2)IL23A(1) IL23A(1) TNFa(2) GS IgG1WT 136/137 VL (SEQ VH (SEQ VL (SEQ VH(SEQ ID NO: 4) ID NO: 7) ID NO: 8) ID NO: 3) Compound BH IL23A(1)TNFa(1) TNFa(1) IL23A(1) VF IgG1WT 138/139 VL (SEQ VH (SEQ VL (SEQ VH(SEQ ID NO: 8) ID NO: 1) ID NO: 2) ID NO: 7) Compound BI IL23A(1)TNFa(1) TNFa(1) IL23A(1) VF IgG4Pro 140/141 VL (SEQ VH (SEQ VL (SEQ VH(SEQ ID NO: 8) ID NO: 1) ID NO: 2) ID NO: 7) Compound BJ IL23A(1)TNFa(1) TNFa(1) IL23A(1) VF IgG4Pro-YTE 142/143 VL (SEQ VH (SEQ VL (SEQVH (SEQ ID NO: 8) ID NO: 1) ID NO: 2) ID NO: 7) TNFa = TNF-alpha, VL =variable domain light chain, VH = variable domain heavy chain, GS =GGGSGGGG (SEQ ID NO: 9), LGGGSG (SEQ ID NO: 10), or both, VF = FNRGES(SEQ ID NO: 11), VEPKSS (SEQ ID NO: 12), or both, IgG1WT = IgG1 wildtype; IgG1KO-YTE = IgG1 with a M252Y/S254T/T256E triple mutation in theFc region and also comprising L234A/L235A mutations, IgG4Pro-YTE = IgG4with a M252Y/S254T/T256E triple mutation in the Fc region and alsocomprising S241P mutation, IgG1KO = truncated Fc region comprisingE234A/E235A mutations, IgG4Pro = comprising S241P mutation. 1^(st) =first polypeptide, 2^(nd) = second polypeptide. The numbering ofmutations is Kabat numbering for a conventional antibody starting withthe antibody convention at CH1.

The below control antibodies were also used for comparison purposes. Thecontrols were monoclonal antibodies that targeted either TNFa or IL23.

TABLE 2B Control compounds VH sequence VL sequence Control antibody 1TNFa(1) TNFa(1) (TNFa monoclonal VH (SEQ VL (SEQ antibody) ID NO: 1) IDNO: 2) Control antibody 2 TNFa(2) TNFa(2) (TNFa monoclonal VH (SEQ VL(SEQ antibody) ID NO: 3) ID NO: 4) Control Antibody 3 IL23A(1) IL23A(1)(IL23 monoclonal VH (SEQ VL (SEQ antibody) ID NO: 7) ID NO: 8)

Example 2. Surface Plasmon Resonance (SPR) Affinity of ExemplaryCompounds

Test compounds were analyzed by SPR to determine affinity for TNF-alphaand IL23A.

Materials and Methods

SPR experiments were performed on a ProteOn XPR36 instrument (Bio Rad).A GLM chip was preconditioned with sequential injections of 60 sec of0.5% SDS, 50 mM NaOH, and 100 mM HCl at a flow rate of 30 μl/min bothvertical and horizontal directions. The preconditioned GLM chip was thenactivated by an injection of EDC (76.7 mg/ml) and sulfo-NHS (21.7 mg/ml)mixture with ratio of 1:1 in 6 horizontal channels. Goat-anti-human IgG(GAHA) Fc gamma (Invitrogen) at a concentration of 30 μg/ml in 10 mM, pH5.0 sodium acetate buffer was immobilized to 8,000 resonance units onthe activated GLM chip in 6 horizontal channels. The chip was finallydeactivated with 1 M ethanolamine HCl in 6 horizontal channels. Theprepared GAHA chip was rotated to vertical direction to capture testcompounds, over 5 vertical channels and the last channel was used as acolumn reference. The captured chip was then rotated again to thehorizontal direction for binding. Linked human IL-23 (BoehringerIngelheim Pharmaceuticals, Inc) with five concentrations, 10.0 nM, 5.00nM, 2.50 nM, 1.25 nM and 0.625 nM, were injected horizontally over thetest compound surfaces for 10 minutes at a flow rate of 40 μl/min in thefollowing running buffer (Bio Rad): phosphate buffer saline (pH 7.4),0.005% Tween 20. The dissociation was allowed for 2 hour. The GAHAsurface was regenerated using short pulse injection (18 seconds) of0.85% phosphoric acid (Bio Rad) at a flow rate of 100 μl/min bothhorizontal and vertical directions after 10 min association and 2 hrdissociation. The regenerated GAHA was ready for another binding cycle.Binding of compounds to human TNF-alpha or cynomologus TNF-alpha wasdone in similar way.

Results

The results in Table 3 show that both compounds tested were able to bindTNF-alpha and IL23 with a dissociation constant (KD) in the picomolarrange.

TABLE 3 KD to KD to human cynomologus KD to human Compound ID TNF-alpha(pM) TNF-alpha (pM) IL23 (pM) Compound A 2.14 7.71 4.28 ± 2.03 CompoundE 4.11 ± 0.68 37.1 ± 16.2 7.00 ± 6.92

Example 3. Flow Cytometry Assessment of Binding to Membrane BoundTNF-Alpha

Test compounds were assessed for their ability to dose dependently bindto cell lines transfected to express membrane bound TNF-alpha.

Material and Methods

All reagents were prepared in flow cytometry staining buffer(BioLegend). Membrane expressed TNF-alpha transfected cell lines (Jurkatand CHO) and parental cell lines were harvested from tissue culturevessels, washed, counted and resuspended to 1×10{circumflex over ( )}6cells/ml in flow cytometry staining buffer. One hundred microliters ofthe cell suspension was added to 96 well microtiter plates and placed onice. Titrations of test compounds were prepared and 50 uL was added tothe cells. After sixty minute incubation on ice, the cell+test compoundswere washed and 50 uL of a secondary antibody (Jackson ImmunoResearch)was added. The samples were incubated in the dark, at 4 C, for 60minutes, followed by washes. After a final wash the cells wereresuspended in 60 uL of fixative (BD Bioscience). Median fluorescencewas determined for each sample in a flow cytometer and plotted versusthe concentration of the test sample. EC₅₀ values were calculated usingthe 4 Parameter Logistic enabled by the Excel add-in XLfit (ActivityBase software, ID Business Solutions, Ltd.). The EC₅₀ values shown beloware Geomeans calculated across multiple experiments for each test sampleand are shown in Table 4.Results

The results shown in Table 4 below demonstrate that the compounds testedbound to membrane bound TNF-alpha in a dose dependent manner.

TABLE 4 EC50 values for membrane bound TNF-alpha. mTNF-Jurkat CellmTNF-CHO Cell Binding EC₅₀ pM Binding EC₅₀ pM Compound ID (Geomean)(Geomean) Compound M 650 950 Compound A 910 890 Compound O 270 770Compound E 200 450 Control antibody 1 310 400 (TNFa) Control antibody 2230 310 (TNFa)

Example 4. In Vitro L929 Cytotoxicity Assay

The compounds were tested for their ability to inhibit TNF-alpha inducedcytotoxicity.

Methods and Materials

This protocol used the PrestoBlue™0 Cell Viability Reagent to determinecytotoxicity of recombinant human TNF-alpha. A more detailed protocolfor the PrestoBlue Cell Viability Protocol can be downloaded from theInvitrogen website (Invitrogen.com). L929 cells were grown andharvested. 1.5×10⁴ cells were transferred to each well of a 96-wellplate for incubated overnight at 37° C. Serial dilutions of compoundswere prepared starting at 5 nM in complete assay medium containing 10μg/ml of actinomycin D and 1000 pg/ml of rhTNF-alpha. The positivecontrols contained 20 ng/ml rhTNF-alpha and 1 μg/ml actinomycin D. Thenegative control contained no TNF-alpha. 10 μL of the dilutions wasadded to corresponding wells and incubated overnight at 37° C. in a 5%CO2. PrestoBlue™ reagent was added to wells and the plate was incubatedfor 2 hour at 37° C. in a 5% CO2. The relative fluorescence unit of eachwell was measured using a Victor™×2 plate reader (excitation: 560 nm,emission: 590 nm). The fluorescent units (Y-axis) versus concentrationof test compound (X-axis) were plotted and the IC₅₀ and IC₉₀ values oftest compounds were calculated by using Graphpad software.

Results

The results in Table 5 show that the tested compounds were able toinhibit TNF-alpha induced cytotoxicity in a dose-dependent manner.

TABLE 5 L929 TNF IC₅₀ Cytotox pM Compound ID IC₉₀ Geomean Compound MIC₅₀ 19 Compound M IC₉₀ 55 Compound A IC₅₀ 20 Compound A IC₉₀ 67Compound N IC₅₀ 34 Compound N IC₉₀ Compound D IC₅₀ Compound D IC₉₀Compound O IC₅₀ 4.2 Compound O IC₉₀ 16 Compound E IC₅₀ 4.1 Compound EIC₉₀ 17 Compound P IC₅₀ 3.4 Compound P IC₉₀ 14 Compound F IC₅₀ 2.5Compound F IC₉₀ 10 Control antibody 1 IC₅₀ 62 (TNFa) Control antibody 1IC₉₀ 230 (TNFa) Control antibody 2 IC₅₀ 20 (TNFa) Control antibody 2IC₉₀ 95 (TNFa)

Example 5. Inhibition of TNF-Alpha Dependent IL-8 Release in HeLa Cells

Anti-TNF test samples were assessed for their ability to inhibit the TNFdependent release of IL8 from the human cell line, HeLa. The sampleswere tested against a high and low concentration of recombinant humanTNF-alpha and a single (high) concentration of recombinant cynomolgusTNF-alpha.Materials and Methods

Briefly, HeLa cells (ATCC) were harvested, washed, counted andresuspended to 4×10{circumflex over ( )}5 cells/ml in a standardcomplete media of (v/v) 10% Fetal Bovine Serum with 1% Penicillin&Streptomycin (CM). One hundred microliters of the HeLa cell suspensionwas added to 96 well microtiter plates. Recombinant human TNF-alpha (R&DSystems) at two concentrations (147 nM or 4.4 nM) as well as generatedrecombinant cynomolgus TNF-alpha (Boehringer Ingelheim Pharmaceuticals,Inc.) (147 nM) were pre-incubated for 30 minutes at 37 C with CM aloneor with titrations of test samples. After the pre-incubation of testsample+TNF-alpha, 100 ul of the mixture(s) was added to the cells andthe test plates were incubated at 37 C with 5% CO₂-humidified air for 20hours. Control samples received either CM (unstimulated controls) orrecombinant TNF-alpha diluted in CM (stimulated controls). After theincubation, supernatants were assayed for IL8 in an ELISA kit (MesoScaleDiscovery) following the manufacturer's instructions. Interpolated IL8pg/ml values were determined for each sample and converted to percent ofcontrol (POC). The POC was plotted versus concentration of the testsample and IC₅₀ and IC₉₀ values were calculated using a 4 ParameterLogistic Model enabled by the Excel add-in XLfit (Activity Basesoftware, ID Business Solutions, Ltd.).

The test compounds were analyzed with respect to the IC₅₀/IC₉₀ asdescribed above, and Geomeans were calculated across multipleexperiments for each test sample and shown in Table 6.

Results

The results in Tables 6 show that the IC₅₀ and IC₉₀ Geomean values forthe tested compounds were similar to the IC₅₀ and IC₉₀ Geomean valuesfor Control Antibody 1 and Control antibody 2. The data demonstratesthat the test compounds dose dependently inhibited the TNF-alpha inducedIL-8 secretion with either human (at two concentrations tested) or cynorecombinant TNF-alpha.

TABLE 6 HeLa IL8 HeLa IL8 HeLa IL8 IC₅₀ Lo-Hu-TNF pM Hi-Hu-TNF pMCyno-TNF pM Compound ID IC₉₀ Geomean Geomean Geomean Compound M IC₅₀ 7.9260 150 Compound M IC₉₀ 48 420 270 Compound A IC₅₀ 8 280 120 Compound AIC₉₀ 41 460 260 Compound N IC₅₀ 9.2 350 170 Compound N IC₉₀ 54 570 330Compound D IC₅₀ 11 380 190 Compound D IC₉₀ 63 590 390 Compound O IC₅₀9.9 430 300 Compound O IC₉₀ 43 760 970 Compound E IC₅₀ 9.2 320 180Compound E IC₉₀ 35 530 600 Compound P IC₅₀ 9.2 410 210 Compound P IC₉₀36 810 810 Compound F IC₅₀ 7.9 350 190 Compound F IC₉₀ 39 660 740Control antibody 1 IC₅₀ 34 330 170 (TNFa) Control antibody 1 IC₉₀ 140490 330 (TNFa) Control antibody 2 IC₅₀ 11 290 280 (TNFa) Controlantibody 2 IC₉₀ 55 520 1200 (TNFa)

Example 6 Inhibition of TNF-Alpha Dependent IL8 in Whole Blood

TNF is a potent inducer of IL8 release from human cells. Compounds weretested for their ability to inhibit TNF-alpha induced IL-8 release inwhole blood samples.

Methods and Materials

Briefly 120 uL of heparinized human whole blood was added to each wellin a 96 well microtiter plate. Assay reagents were prepared in astandard T cell media (TCM). Titrations of test samples were prepared at10× concentrations and pre-incubated with a 10× concentration of humanrecombinant TNF (100 ng/ml, R&D Systems) for 1 hour at 37 C. After thispre-incubation, 30 ul of the cytokine/test compound mixture was added tothe whole blood along with 30 uL of appropriate controls in TCM andincubated at 37 C with 5% CO₂-humidified air for 48 hours. Controlsamples received either TCM (unstimulated controls) or recombinant humanTNF-alpha diluted in TCM (stimulated controls). After the incubation,supernatants were assayed for IL8 in an ELISA kit (MesoScale Discovery)following manufacturer's instructions. Interpolated IL8 pg/ml valueswere determined for each sample and converted to percent of control(POC). The POC was plotted versus concentration of the test sample andIC₅₀ and IC₉₀ values were calculated using a 4 Parameter Logistic Modelenabled by the Excel add-in XLfit (Activity Base software, ID BusinessSolutions, Ltd.).

The test compounds were analyzed with respect to the IC₅₀/IC₉₀ asdescribed above, and Geomeans were calculated across multipleexperiments for each test sample and shown in Table 7.

Results

The results in Table 7 show that the IC₅₀ and IC₉₀ Geomean values forthe tested compounds were similar to the IC₅₀ and IC₉₀ Geomean valuesfor Control antibody 1 and control antibody 2. The data demonstratesthat the test compounds dose dependently inhibited the TNF-alpha inducedIL8 release in human whole blood.

TABLE 7 TNF-IL8 IC₅₀ Whole Blood pM Compound ID IC₉₀ Geomean Compound MIC₅₀ 380 Compound M IC₉₀ 790 Compound A IC₅₀ 360 Compound A IC₉₀ 490Compound N IC₅₀ 270 Compound N IC₉₀ 520 Compound D IC₅₀ 560 Compound DIC₉₀ 1100 Compound O IC₅₀ 320 Compound O IC₉₀ 470 Compound E IC₅₀ 340Compound E IC₉₀ 610 Compound P IC₅₀ 290 Compound P IC₉₀ 420 Compound FIC₅₀ 310 Compound F IC₉₀ 450 Control antibody 1 IC₅₀ 320 (TNFa) Controlantibody 1 IC₉₀ 490 (TNFa) Control antibody 2 IC₅₀ 330 (TNFa) Controlantibody 2 IC₉₀ 600 (TNFa)

Example 7. NF-kappaB and STAT3 Phosphorylation Assays

IL23 engagement with its heterodimeric receptor complex (IL12Rβ1-IL23R)results in the downstream phosphorylation of Signal transducer andactivator of transcription 3 (STAT3). TNF engagement with its receptors(TNFR1/TNFR2) results in the downstream phosphorylation of nuclearfactor of kappa light polypeptide gene enhancer in B-cells (NF-κB).Compounds were assessed for their ability to inhibit TNF-dependentphosphorylation of NF-κB in Jurkat cells, and IL23-dependentphosphorylation of STAT3 in DB cells.

Methods and Materials:

Briefly, cultures of Jurkat cells (ATCC) and DB cells (ATCC) growing inlog phase were harvested, washed, counted and resuspended to2×10{circumflex over ( )}7 cells/mL in a standard complete media (CM;RPMI1640 with (v/v) 10% FCS and 1× Penicillin-Streptomycin(Invitrogen)). Titrations of test samples were prepared at 4×concentrations and pre-incubated with a mixture of 4× human recombinant1L23 (Boehringer Ingelheim Pharmaceuticals, Inc.) and recombinant humanTNF (R&D Systems) for 1 hour at 37 C. After the pre-incubation of thetest reagent+cytokine mixture, 100 μL of the mixture was added to wellscontaining 100 μL of cells in duplicate. Controls were setup as follows:100 μL of the diluted TNF/IL23+100 μL combined cells (stimulatedcontrol), or 100 μL of CM+100 μL combined cells (unstimulated control).The assay plates were incubated for exactly 10 minutes at 37° C. with 5%CO₂-humidified air. After the incubation, cell lysates were prepared andp-NF-κB and p-STAT3 was assessed following the manufacturer'sinstructions (MesoScale Discovery). p-NF-κB and p-STAT-3 raw values weredetermined for each sample and converted to percent of control (POC).The POC was plotted (Y-axis) versus concentration of the test agent(X-axis). IC₅₀ and IC₉₀ values were calculated using the 4 ParameterLogistic Model enabled by the Excel add-in XLfit (Activity Basesoftware, ID Business Solutions, Ltd.).

The test compounds were analyzed with respect to the IC₅₀/IC₉₀ asdescribed above, and Geomeans were calculated across multipleexperiments for each test sample and shown in Table 10. Note: this assayprovides confidence that that the dual molecule is capable ofneutralizing both downstream signaling events. The assay time point isoptimal for the p-NF-κB signal only and therefore the calculatedIC₅₀/IC₉₀ does not reflect the overall potencies in a quantitativemanner.

Results

The results in Table 8 show that the test compounds were able to inhibitboth TNF-alpha induced NF-kB phosphorylation as well as IL23 inducedphosphorylation of STAT3 in DB cells.

TABLE 8 Dual Phospho Dual Phospho IC₅₀ Jurkat-pNf-Kb pM DB-pSTAT3 pMCompound ID IC₉₀ Geomean* Geomean* Compound M IC₅₀ 290 190 Compound MIC₉₀ 680 580 Compound A IC₅₀ 300 200 Compound A IC₉₀ 480 500 Compound NIC₅₀ 300 210 Compound N IC₉₀ 620 760 Compound D IC₅₀ 270 170 Compound DIC₉₀ 810 560 Compound O IC₅₀ 210 210 Compound O IC₉₀ 740 580 Compound EIC₅₀ 260 230 Compound E IC₉₀ 340 770 Compound P IC₅₀ 290 340 Compound PIC₉₀ 340 630 Compound F IC₅₀ 280 360 Compound F IC₉₀ 760 980 Controlantibody 1 IC₅₀ 360 NA (TNFa) Control antibody 1 IC₉₀ 660 NA (TNFa)Control antibody 2 IC₅₀ 260 NA (TNFa) Control antibody 2 IC₉₀ 420 NA(TNFa) Control antibody IC₅₀ NA  89 3(IL23A) Control antibody IC₉₀ NA230 3(IL23A) *Results are semi-quantitative and optimized more to theTNF readout. NA; No Activity

Example 8 Inhibition of IL23 Induced STAT3 Phosphorylation in DB Cells

IL23 engagement with its heterodimeric receptor complex (IL12Rβ1-IL23R)results in the downstream phosphorylation of Signal transducer andactivator of transcription 3 (STAT3). Anti-IL23 test samples wereassessed for their ability to inhibit the IL23 dependent phosphorylationin the human DB cell line.

Materials and Methods:

Briefly 100 uL of the human DB cell line (ATCC) grown in log phase wasadded to each well in a 96 well microtiter plate at a concentration of1×10{circumflex over ( )}7 cells/ml. Assay reagents were prepared in acomplete media (CM; RPMI1640 with (v/v) 10% Fetal Calf Serum and 1×Penicillin-Streptomycin (Invitrogen)). Titrations of test samples wereprepared at 4× concentrations and pre-incubated with a 4× concentrationof human recombinant IL23 (Boehringer Ingelheim Pharmaceuticals, Inc.)for 1 hour at 37 C. After this pre-incubation, 100 ul of thecytokine/test sample mixture was added to the 100 uL of DB cells andincubated at 37 C with 5% CO₂-humidified air for 30 minutes. Controlsamples received either CM (unstimulated controls) or recombinant humanIL23 diluted in CM (stimulated controls). After the incubation, celllysates were prepared and pSTAT3 was assessed following themanufacturer's instructions (MesoScale Discovery). Raw pSTAT3 valueswere determined for each sample and converted to percent of control(POC). The POC was plotted versus concentration of the test sample andIC₅₀ and IC₉₀ values were calculated using a 4 Parameter Logistic Modelenabled by the Excel add-in XLfit (Activity Base software, ID BusinessSolutions, Ltd.). The test compounds were analyzed with respect to theIC₅₀/IC₉₀ as described above, and Geomeans were calculated acrossmultiple experiments for each test sample and shown in Table 9.

Results

The results in Table 9 show that the IC₅₀ and IC₉₀ Geomean values forthe tested compounds were similar to the IC₅₀ and IC₉₀ Geomean valuesfor an anti-IL23Ap19 control antibody. The data demonstrates that thetest compounds dose dependently inhibited the IL23 inducedphosphorylation of STAT3 in DB cells.

TABLE 9 hIL23 pSTAT3 IC₅₀ DB assay pM Compound ID IC₉₀ Geomean CompoundM IC₅₀ 190 Compound M IC₉₀ 530 Compound A IC₅₀ 210 Compound A IC₉₀ 420Compound N IC₅₀ 240 Compound N IC₉₀ 510 Compound D IC₅₀ 280 Compound DIC₉₀ 560 Compound O IC₅₀ 300 Compound O IC₉₀ 720 Compound E IC₅₀ 300Compound E IC₉₀ 700 Compound P IC₅₀ 300 Compound P IC₉₀ 620 Compound FIC₅₀ 260 Compound F IC₉₀ 600 Control antibody IC₅₀ 160 3(IL23A) Controlantibody IC₉₀ 310 3(IL23A)

Example 9. Human IL-23 Dependent Mouse Splenocyte Assay (MSA)

A mouse splenocyte based assay was used to assess the ability ofanti-human IL23 test samples to inhibit the induction of mouse IL17 byhuman recombinant IL23 and recombinant cynomolgus IL23 in mousesplenocyte cultures.

Materials and Methods:

Briefly, mononuclear cells from mouse spleens (female C57BL/6 less than13 weeks of age; JAX) were isolated washed, counted and resuspended to4×10{circumflex over ( )}6 cells/ml in a standard T cell media (TCM).One hundred microliters of the mIL2/splenocyte suspension was added to96 well microtiter plates. Recombinant human IL23 (Boehringer IngelheimPharmaceuticals, Inc.) or recombinant cynomolgus IL23 (BoehringerIngelheim Pharmaceuticals, Inc.) was diluted in TCM and pre-incubatedfor 2 hours at 37 C with TCM alone or with titrations of test samples.After the pre-incubation of test sample+IL23, 100 ul of the mixture wasadded to the cells and the test plates were incubated at 37 C with 5%CO₂-humidified air for 48 hr. Control samples received either TCM(unstimulated controls) or recombinant human IL23 diluted in TCM(stimulated controls) After the incubation, mouse IL17 levels weredetermined from the supernatant using the Quantikine® Mouse IL-17Immunoassay according to the manufacturer's instructions (R&D Systems).Interpolated mIL17 pg/ml values were determined for each sample andconverted to percent of control (POC). The POC was plotted versusconcentration of the test sample and IC₅₀ and IC₉₀ values werecalculated using a 4 Parameter Logistic Model enabled by the Exceladd-in XLfit (Activity Base software, ID Business Solutions, Ltd.). Theanti-IL23 test samples were analyzed with respect to the IC₅₀/IC₉₀ asdescribed above and Geomeans were calculated across multiple experimentsfor each test sample and shown in Table 10.

Results

The results in Table 10 show that the tested compounds were able toinhibit both human and cynomolgus-IL23 induced mouse splenocyte releaseof IL17.

TABLE 10 huIL23 CynoIL23 IC₅₀ MSA pM MSA pM Compound ID IC₉₀ GeomeanGeomean Compound M IC₅₀ 140 120 Compound M IC₉₀ 1600 890 Compound A IC₅₀180 120 Compound A IC₉₀ 1700 730 Compound N IC₅₀ 230 140 Compound N IC₉₀2000 940 Compound D IC₅₀ 190 160 Compound D IC₉₀ 2200 1100 Compound OIC₅₀ 210 120 Compound O IC₉₀ 2200 800 Compound E IC₅₀ 200 77 Compound EIC₉₀ 1700 1200 Compound P IC₅₀ 200 97 Compound P IC₉₀ 1400 1500 CompoundF IC₅₀ 170 69 Compound F IC₉₀ 2300 1500 Control antibody IC₅₀ 53 173(IL23A) Control antibody IC₉₀ 350 240 3(IL23A)

Example 10 Inhibition of IL23 Induced Phosphorylation of STAT3

IL23 engagement with its heterodimeric receptor complex (IL12R131-IL23R)results in the downstream phosphorylation of Signal transducer andactivator of transcription 3 (STAT3). Compounds were tested for theability to inhibit IL23 induced STAT3 activation in DB stabletransfected cells

Materials and Methods

The cells were stimulated with a final concentration of 15 ng/ml of IL23protein. This dose was estimated to be the EC60 according to previousexperiments, while allowing for inhibition with the tested compound.Cells were plated, compound dosed, and IL-23 added (in that order) andincubated overnight. If the compound inhibited cell stimulation, STAT3was downregulated, leading to less luciferase activity.

Results

The results in Table 11 show that the tested compounds were able toinhibit IL23 induced phosphorylation of STAT3.

TABLE 11 IL23A pSTAT3 IC₅₀ (MG) pM Compound ID IC₉₀ Geomean Compound MIC₅₀ 120 Compound M IC₉₀ 300 Compound A IC₅₀ 130 Compound A IC₉₀ 1100Compound O IC₅₀ 160 Compound O IC₉₀ 650 Compound E IC₅₀ 140 Compound EIC₉₀ 830 Compound P IC₅₀ 69 Compound P IC₉₀ 480 Compound F IC₅₀ 90Compound F IC₉₀ 650 Control antibody IC₅₀ 35 3(IL23A) Control antibodyIC₉₀ 140 3(IL23A)

Example 11. Further IL23-A Stat3 Assays

Further experiments were run similarly to Example 8 to test forinhibition of IL23 induced activation of STAT3.

Methods and Materials

DB-STAT3Luc10 Clone 10 suspension cells were grown in RPMI1640+10% FBS.20,000 cells were added per well of 96 well plates at 80 ul/well of cellsuspension. 10 ul of one of the serially diluted test compounds wasadded to each well. 15 ng/mL of recombinant human IL-23 was added toeach well, with certain wells contained only test compounds and noIL-23, for comparison. The plates were incubated overnight at 37° C./5%CO2. Luciferase activity was assayed using Steady-Glo (Promega andOne-Glo (Promega and the results were read on Envision Reader.

Results

The IC₅₀ and IC₉₀ for the tested compounds are shown in Table 12 andTable 13. These tables show that the compounds inhibited IL-23-dependentSTAT3 activation in a dose dependent manner.

TABLE 12 IC₅₀ IC₉₀ Compound ID (pM) (pM) Compound N 235.2 873.7 Controlantibody 96.5 192.6 3(IL23A) Compound D 185.6 871.6 Compound E 211.9965.1 Control antibody 104.3 198.8 3(IL23A) Compound G 220.2 1151.0Compound C 162.7 620.4 Control antibody 80.3 181.3 3(IL23A)

TABLE 13 First test Second Test GEOMEAN IC₅₀ IC₉₀ IC₅₀ IC₉₀ IC₅₀ IC₉₀Compound ID (pM) (pM) (pM) (pM) (pM) (pM) Control antibody 96.5 192.693.1 190.8 3(IL23A) 104.3 198.8 80.3 181.3 Compound N 178.6 856.4 235.2873.7 205.0 865.0 Compound D 178.1 657.2 185.6 871.6 181.8 756.8Compound E 170.2 764.9 211.9 965.1 189.9 859.2 Compound G 187.1 600.5220.2 1151.0 203.0 831.4 Compound C 134.9 352.6 162.7 620.4 148.1 467.7

Example 12. Inhibition of Mouse IL17A and IL22 Release Induced by HumanRecombinant IL23

Test compounds were assessed for their ability to inhibit human IL23induced cytokine release in C57/Bl6 mice. IL17A and IL22 secretion aremeasured after intradermal injection of IL23

Materials and Methods

Briefly, C57BL/6 female mice (7-10 weeks old, Charles River) wererandomly divided into 8 groups, 8 animals/group and given a 100 μlintraperiotoneal injection of either citrate buffer (20 mM NaCitrate,115 mM NaCl, pH 6.0) or test compounds at equivalent molar dose of 1.3,0.4 and 0.13 mg/kg vs. 1, 3 and 0.1 mg/kg respectively.

One hour after test compound dosing mice were anesthetized viaisoflurane (Butler Schein) and given a 20 μl intradermal injection ofeither 0.1% BSA (Sigma) control or 15 μg/ml (0.3n) rhIL23 (generatedin-house) diluted in saline (Invitrogen) to both ears. Intradermalchallenges were repeated daily for 2 consecutive days. Twenty-four hoursafter the second challenge the mice were sacrificed via cervicaldislocation and each ear was removed. Ear tissue was homogenized in 1 mlof homogenization buffer (HBSS (Gibco); 0.4% Triton X-100 (Sigma); 1×SigmaFast Protease Inhibitor (Sigma)) using a MP Biomedicals Fast-Prep24 homogenizer. Homogenized samples are centrifuged at 4 C for 10 minand supernatant collected. Supernatants were assayed for the presence ofmouse IL17A and IL22, using the Quantikine® Mouse IL-17 and mouse IL-22Immunoassays according to the manufacturer's instructions (R&D Systems).Interpolated cytokine pg/ml values were determined for each sample. Themean pg/ml levels for each treatment group were determined andsignificance compared to control calculated using the One-way ANOVAfollowed by Dunnett's multiple comparisons test. Results are shown inFIG. 4 .

Results

The results in FIG. 4 show that treatment with a single intraperitonealdose of test compound was able to significantly inhibit the release ofmouse IL17 and IL22 in the skin induced by two daily consecutive intradermal injections of recombinant human IL23.

Example 13 Inhibition of Exogenous Human TNF-Alpha Dependent CytokineRelease in C57/Bl6 Mice

Test compounds were assessed for their ability to inhibit human TNFinduced cytokine release in C57/Bl6 mice after exogenous exposure tohuman TNF. Serum KC and IL-6 secretion are measured followingintraperitoneal administration of human TNF.

Materials and Methods

Briefly, C57BL/6 female mice (8-9 weeks old, Jackson Labs) were randomlydivided into 8 groups, 8 animals/group and given a 200 μlintraperiotoneal injection of either phosphate buffered saline (Sigma)or test compound at equivalent molar dose of 13.3, 4 and 1.3 mg/kg vs.10, 3 and 1 mg/kg respectively.

Two hour after test compound dosing mice were anesthetized viaisoflurane (Butler Schein) and given a 200 μl intraperitoneal injectionof either 0.1% BSA control or 15 μg/ml (3 μg) rhTNF (R&D Systems)diluted in saline (Sigma). Two hours after the TNF challenge the micewere anesthetized via isoflurane, whole blood was collected and micewere then sacrificed via cervical dislocation. Whole blood wascentrifuged at 12,000 rpm for 10 minutes and plasma collected. Plasmawas assayed for the presence of mouse KC and IL-6, using the MultiPlex®Mouse KC and mouse IL-6 Immunoassays according to the manufacturer'sinstructions (MSD). Interpolated cytokine pg/ml values were determinedfor each sample. The mean pg/ml levels for each treatment group weredetermined and significance compared to control calculated using theOne-way ANOVA followed by Dunnett's multiple comparisons test. Resultsare shown in Figure X.

Results

The results in FIG. 5 show that treatment with a single intraperitonealdose of test compound was able to significantly inhibit the release ofmouse KC and IL-6 in serum by intraperitoneal injection of recombinanthuman TNF.

Example 14. Pharmacokinetics of Compounds in Cynomolgus Monkeys

Materials and Methods

Single intravenous (IV) dose PK studies for two pairs of compounds(Compound M and Compound A; and Compound O and Compound E) wereconducted in male cynomolgus monkeys (N=3 per group) naïve to biologics,and conducted according to the guidelines of Institutional Animal Careand Use Committee. IV doses were administered at 1 mg/kg as 10 min IVinfusion. Serum samples were collected at pre-dose, 1, 4, 8 hr on theday of dosing, and 1, 2, 3, 4, 5, 7, 10, 14, 21, 28, 35, and 42 (1008hr) days post dosing for Compound M and Compound A; and only up to Day14 for Compound 0 and Compound E. Serum concentrations of the dosedmolecules were measured by a ligand binding assay (ELISA).

Calibration standard curve and quality control (QC) samples wereprepared in 100% serum for each analyte. Each standard curve consistedof seven non-zero points starting at 10240 ng/mL then serially diluted3×. A blank sample (matrix without analyte) was also included. Four QCsamples at low, medium, and high ranges were prepared starting at 2560ng/mL then serially diluted four-fold. The standard curve and QC sampleswere stored frozen until sample analysis at which time they were diluted20 times to mimic study samples. The standard curve and QC samples wereincluded in duplicate during each analytical run. The lower and upperlimits of quantification were defined as the lowest and highest standardcurve points to reproducible have a back-calculated concentration thatdoes not exceed 25 percent (%) of the nominal concentration. Theacceptance criterion for the standard curve points and QC samples was 25percent (%) of the nominal concentration.

Nunc ELISA plates were coated with 1 μL of monkey adsorbed goatanti-human IgG (Southern Biotech) as the capture reagent and incubatedovernight at 2-8° C. After washing and blocking the plates with the washbuffer (0.05% (v/v) Tween 20 in phosphate buffered saline (PBS)) andblocking buffer (5% bovine serum albumin (BSA) in PBS), standard, QC,and unknown samples, diluted 1:20, 1:400 and 1:8000 with 5% monkey serum(monkey serum from Innovative Research) were added to the plate wellsand incubated for 1 hour at room temperature. The plate wells werewashed with the washing buffer and added with monkey adsorbedbiotinylated goat anti-human IgG (Southern Biotech) as the secondaryreagent and incubated at room temperature for 1 hour. The plates werewashed 3 times and added with 100 μL of 1 μg/mL peroxidase-conjugatedstreptavidin for 15 min at room temperature, followed by further 3 timeswashing and the addition of 100 μL of 3,3′,5,5′-Tetramethylbenzidine(TMB, BioFX) substrate for 3-4 min at room temperature. The reaction wasstopped by adding 100 μL of stop solution (BioFX) and the absorbance wasmeasured using Molecular Devices plate reader with SoftmaxPro software,version 5.4.1.

Results

Single IV dose PK studies for two pairs of test compounds (Compound Mand Compound A; and Compound 0 and Compound E) were conducted in malecynomolgus monkeys (N=3 per group) naïve to biologics. The testcompounds were dosed at 1 mg/kg as 10 min IV infusion. Serum sampleswere collected at pre-dose, 1, 4, 8 hr on the day of dosing, and 1, 2,3, 4, 5, 7, 10, 14, 21, 28, 35, and 42 (1008 hr) days post dosing forCompound M and Compound A; and only up to Day 14 for Compound 0 andCompound E. Serum concentrations of the dosed molecules were measured bya ligand binding assay (ELISA).

Serum concentrations (mean and SD) for each of the molecules aresummarized in Table 14.

TABLE 14 Serum concentrations (mean ± SD, N = 3) for the test compoundsin cynomolgus monkey Compound M Compound A Compound O Compound E TimeMean SD Mean SD Mean SD Mean SD (day) (nM) (nM) (nM) (nM) (nM) (nM) (nM)(nM) 0 0 0 0 0 0 0 0 0 0.042 89.62 26.41 115.60 15.44 96.69 23.15 107.5710.43 0.167 75.30 17.97 108.03 22.92 92.29 19.75 102.36 4.81 0.333 69.719.19 89.15 16.32 65.27 11.53 86.45 10.65 1 54.49 13.94 63.68 6.69 30.1012.41 52.87 8.85 2 35.17 8.93 49.95 6.04 7.44 3.22 32.51 6.30 3 18.586.66 43.51 6.64 4.14 0.70 24.65 6.49 4 13.86 5.61 40.02 8.50 2.76 0.2519.02 4.84 5 9.55 3.40 43.69 19.63 1.96 0.09 13.98 3.84 7 3.76 1.3127.84 3.29 1.08 0.21 8.49 3.38 10 BQL BQL BQL BQL BQL BQL 0.3797 NA 14BQL BQL BQL BQL BQL BQL BQL BQL 21 BQL BQL BQL BQL 28 BQL BQL BQL BQL 35BQL BQL BQL BQL 42 BQL BQL BQL BQL BQL: below quantitation limit NC: notcalculated, N = 1

Pharmacokinetic (PK) parameters of these test compounds were calculatedusing the software Phoenix WinNonlin 6.1 (Certara, Md., USA) usingnon-compartmental approach for IV infusion dose. Serum samples thatshowed precipitous drop in the concentrations at any time point afterdosing and all subsequent samples in that particular animal wereexcluded from the PK parameter estimation. Additional analysis showedthat this sudden drop in the concentrations after the first few days wasdue to the development of anti-compound antibodies for a humanizedbiologic molecule in monkey. Only the first seven day data fromindividual animals were included in the PK analysis. Concentration-timeplots are shown in FIGS. 2 and 3 for the two pairs of test compounds.Key PK parameters (mean±SD) for the two pairs of test compounds aresummarized in Table 15.

TABLE 15 Key PK parameters (mean ± SD; N = 3) of the two test compoundpairs in cynomolgus monkey after 1 mg/kg IV 10 min infusion dose AUC CLVss T½ MRT Compound ID (nM · day) (mL/day/kg) (mL/kg) (day) (day)Compound M 186 ± 48.3 28.2 ± 7.8 62.4 ± 13.7 1.6 ± 0.04 2.2 ± 0.1Compound A 592 ± 68.3  8.5 ± 1.1 71.5 ± 8.9  6.2 ± 0.5 8.4 ± 1.0Compound O 98.0 ± 14.7  51.8 ± 7.8 76.1 ± 30.3 2.3 ± 0.8 1.4 ± 0.4Compound E 244 ± 54.6 21.2 ± 4.5 65.0 ± 3.9  2.5 ± 0.3 3.1 ± 0.5

Compound A, the test compound with YTE mutation showed a 3.3-foldreduction in clearance (CL) and a 3.9-fold increase in terminalhalf-life (T½) compared to the corresponding test compound notcontaining YTE mutation (Compound M). Compound E, the test compound withYTE mutation showed a 2.4-fold reduction in CL compared to thecorresponding test compound not containing YTE mutation (Compound O).

Example 15. Predicting Human PK and Human Dose of Exemplary Compound

Predicting Human PK:

Human PK prediction of Compound E was done by allometric scaling fromthe PK parameters obtained in cynomolgus monkey using a factor of 2-foldreduction in clearance in humans compared to monkey while maintainingsame volume of distribution. Thus, predicted clearance in humans is 12.1mL/day/kg with a terminal half-life of 7.4 days.

Predicting Human Dose:

Human dose prediction was done based on extensive exposure-efficacy dataavailable from clinical trials of golimumab in diverse patientpopulations. Golimumab (Simponi®) is approved to treat rheumatoidarthritis (RA), ankylosing spondylitis (AS), and psoriatic arthritis(PsA) patients with 50 mg monthly subcutaneous (SC) doses, andulcerative colitis (UC) patients with 100 mg monthly SC doses. Simponi®achieves a Ctrough of approximately 3.2 nM in RA patients (50 mg monthlySC doses) and 9.7 nM (100 mg monthly SC doses) in UC patients(SimponiER) BLA, 2009; Sandborn, 2013). These are used as benchmarks fortherapeutic Ctroughs for AS and CD, respectively. Ctrough levels at theclinically approved doses of Stelara are about 6 nM. Based on theobservation of a 3-fold higher potency of Compound Ecompared toustekinumab (Stelara®) Ctrough values of ˜2 nM are needed for CompoundEto cover IL23. As the Ctrough concentration for covering TNF is greaterthan that for covering IL23, the 9.7 nM Ctrough for Simponi® was usedfor dose projections.

Compartmental modeling of PK data in the cynomolgus monkey(2-compartment model) followed by Monte-Carlo simulations using a 2-foldreduction scaling of CL, 73% bioavailability, and simultaneously varyingclearance and distributional rate constants with a nominal 30% CV andlog-normal distribution shows that 54 mg (90% confidence intervals 31-90mg) SC doses administered every 2 weeks will maintain a Ctrough of 9.7nM.

Example 16. Purification of Compounds

Methods

Compounds were purified using Mab Select SuRe as an affinitypurification step. High salt washes are avoided in order to preventaggregation. Elution was performed using Sodium Acetate buffer pH 3.5.Following Mab Select SuRE purification the sample was neutralized andapplied to a Hydroxyapatite Type I resin and eluted using variousconcentrations of phosphate buffer. Monomer peak elutes ˜140 mMNaPhosphate 100 mM NaCl pH 7.0 and aggregate peak eluted at −200 mMNaPhosphate 100 mM NaCl pH 7.0. Following hydroxyapatite, the sample wasconsistently >95% monomer.

Sedimentation velocity (SV) experiment via Analyticalultracentrifugation (AUC) was used to provide information on samplepurity and aggregation states. Samples were centrifuged in an optimaXL-I (Beckman Coulter, Fullerton, Calif.) at 20° C. using an An60Tifour-hole rotor running at 40,000 rpm. The sedimentation process wasmonitored by ultraviolet absorbance at 280 nm, using correspondingdilution buffer as reference buffer. The variation in the concentrationdistribution in the ultracentrifuge cell with time was collected usingXL-I operating software and was analyzed using the continuous c(S)distribution model in the SEDFIT software (version 14.1) to give thedistribution of sedimentation coefficient. Monomer percentage wascalculated based on the integrated peak area.

Results

The results of purification of the compounds are shown in Table 16. Thedata show that the compounds have high purity and homogeneity indicatinggood stability.

TABLE 16 Parameter Compound A Compound E Percent monomer 99.4 99.0(sedimentation velocity)

Example 17: Mass Spectrometry Profile of Compounds

Methods

Native Sample

This procedure yielded the intact mass of the compound or protein. 2 ulof sample was injected onto an Agilent PoroShell 300SB-C8 column, Sum,(75×1.0 mm). The column temperature was 80° C. and flow rate was 50ul/min. The compound or protein was eluted off the column with agradient from 20% B at 0 minutes to 85% B at 10 minutes. Mobile phase Awas Water/Acetonitrile/Formic Acid (99/1/0.1) and Mobile phase B wasAcetonitrile/Water/Formic Acid (95/5/0.1). The effluent was directed toan Agilent 6210 TOF mass spectrometer, which was scanned from mass 600to mass 3200. The raw data was deconvoluted with the program MassHunter.

Reduced Sample

This procedure yielded the mass of the protein or the light chain andthe mass of the heavy chain. 2 ul of 50 mM TCEP was added to 10 ul ofsample and 10 ul of 8M Guanidine and incubated for 15 minutes at 37° C.2 ul of this sample was injected as above, with the followingdifferences: the column temperature was 60° C. and the mass range was600-2000.

Deglycosylated Sample

This procedure yielded the deglycosylated mass of the protein or thelight chain and the heavy chain. 10 ul of sample, 10 ul of 200 mMNH₄HCO₃, 2 ul 50 mM TCEP, and 1 ul (1:10) PNGase F (or 1 uL QAdeglycosylation mix if O-linked glycosylations were present) wereincubated for 3 hours at 37° C. The incubation was increased toovernight for heavily glycosylated samples. Then, 25 ul 8M Guanidine and4 ul of 50 mM TCEP were added and incubated for 15 minutes at 37° C.This sample was injected as above for reduced sample.

Protein Peptide Mapping by Mass Spectrometry

25 ul of sample was added to 25 ul of 8M urea in 400 mM ammoniumbicarbonate. 5 ul of 50 mM TCEP was then added and the sample wasincubated for 15 minutes at 60° C. After cooling the sample to roomtemperature, 5 ul of 150 mM iodoacetamide was added and the sample wasincubated at room temperature for 15 minutes. After adding 40 ul ofwater, 5 ul of trypsin in 1 mM HCl was added to give a final enzyme:substrate ratio of 1:50. The sample was incubated at 37° C. overnight. 5ul was then injected onto a Thermo Hypurity C18 column, 100×1.0 mm. Flowrate was 80 ul/min. The protein was eluted off the column with agradient from 0% B at 0 minutes to 40% B at 33 minutes. Mobile phase Awas Water/Acetonitrile/Formic Acid (99/1/0.1) and Mobile phase B wasAcetonitrile/Water/Formic Acid (95/5/0.1). The effluent was directed toa Thermo Orbitrap Velos mass spectrometer. The first scan event was inthe FT, and scans from mass 300 to mass 2000 with a resolution of30,000. The second through the seventh scan events were in the IT (iontrap) and fragmented the 6 most intense ions from the first scan event.Peptides containing glycosylation were profiled by manual extraction andpercentages calculated based on peak heights.

Results

The results are shown in Table 17. The data indicate the intended aminoacid sequence and structure has been expressed and recovered withoutunexpected heterogeneity. The glycosylation pattern is typical of aconventional antibody expressed in CHO cells and does not show anyatypical structures.

TABLE 17 Parameter Compound A Compound E Mass Spectrometry:Intact/Matches Intact/Matches Intact Molecular Weight Profile SequenceSequence Mass Spectrometry: Not Determined Similar to CHO GlycosylationProfile expressed IgG

Example 18: Thermal Stability of Compounds

Methods

Thermal unfolding and aggregation of 2 mg/ml solutions of the compoundsin phosphate buffer were monitored from 20° C. to 110° C. at a scan rateof 60° C./hr via an automated capillary DSC (MicroCal, LLC, Boston). Twoscans with the corresponding buffer were performed to establishinstrument thermal history and to obtain the instrument baseline foreach sample, with the average of these scans subtracted from thesubsequent protein thermogram to obtain the apparent heat capacity.Normalized scans were then analyzed with Origin 7.0. Pre-transitionbaselines were subtracted from each resulting heat capacity thermogram,to give the resulting excess heat capacity (Cp,ex) as a function oftemperature. Reported values of transition temperatures (Tm) representpositions of peak maxima determined by visual inspection of theexperimental thermograms.

Results

The results are shown in Table 18. The data show that the compounds arestable and would predict the ability to have a long shelf-life.

TABLE 18 Parameter Compound A Compound E Thermal Stability (° C.) 57.9,72.1, 82.9 67.6, 83.1

Example 19: Solubility of Compounds

Methods

The compound samples were concentrated gradually to a concentration ashigh as possible without precipitation observed using Amicon Ultracentrifugal filter with cut-off molecular weight of 50,000 Dalton(Millipore, Billerica). The concentrated protein solutions were thenanalyzed in SV experiment via AUC to provide information on samplepurity and aggregation states (refer to Example 16 regardingpurification for method details).

Results

The results are shown in Table 19. The data show that the compounds aresoluble and stable retaining a high percentage of monomer withoutformulation or added excipients.

TABLE 19 Parameter Compound A Compound E Solubility (Concentration) 48mg/ml 51 mg/ml Percent Monomer 98.6 97.0

Example 20: Valence of Compounds

Methods

The valence measurement for the compound samples in 50 mM KCl and 10 mMsodium acetate buffer at pH 5.0 was performed on a Beckman Coulter(Fullerton, Calif.) ProteomeLab PA800™ apparatus equipped with anultraviolet (UV) absorbance detector, with a working wavelength of 214nm. The system was maintained at 20° C. and an eCap amine capillary withan inner diameter of 50 μm (Beckman Coulter, part #477431) was used. Thecapillary was rinsed with 100 mM NaOH, amine regeneration solution(Beckman Coulter, part #477433) and running buffer before each sampleinjection. Migration times for the samples were measured at voltages of10 kV, 14 kV, and 18 kV. Dimethylformamide (DMF) (0.005%) (Pierce) wasused as an electroosmotic flow (EOF) marker. Data were acquired using 32Karat™ software (v7.0). Diffusion coefficient was determined from SVexperiment via AUC.

Results

The valence data (see Table 20) indicate colloidal stability of thecompounds in solution, i.e. net interaction of protein and protein insolution. The compounds with valence greater than 15 have strong netrepulsive interaction and high potential to be formulated at highconcentration.

TABLE 20 Parameter Compound A Compound E Valence, pH 5.0 20.9 24.4

Example 21: Predicted In Silico Immunogenicity

Methods

Immunogenicity of protein therapeutics was predicted in silico byutilizing a computational tool, EpiMatrix that was developed by EpiVax,Inc. (Providence, R. I.). EpiMatrix incorporates the prediction ofT-helper epitope as well as the T-regitope, of which the former is toprovoke an immune response while the latter is inhibitory. Briefly, theprotein sequence was first parsed into overlapping 9-mer peptide framesthat has been proven the core of class II HLA binding. The bindingpotential of 9-mer peptides to each of eight common class II HLA allelesare evaluated based on experimental data or computational prediction. Ascore is generated to reflect the binding potential of the 9-mer peptideto each HLA allele and normalization is performed to make it possible tocompare any 9-mer across multiple HLA alleles and enable immunogenicityprediction on a global scale. In the end the program generates anoverall ‘immunogenicity score’, tReg Adjusted Epx Score, that togetherwith other immunogenicity determinants helps to make an informeddecision of the likelihood that the compounds will provoke an immuneresponse in vivo.

Results

The results are shown in Table 21. The overall immunogenicity scores forthese compounds are low and predict that these compounds are not likelyto illicit a strong immune response in vivo.

TABLE 21 Parameter Compound A Compound E EpiVax −37.7, −35.6 −31.1,−46.8 Normalization of allele-specific scores

Example 22: Whole Blood Stability of Compounds

Methods

A whole blood interference assay was developed on an Octet RED96 todetect the effects of non-specific binding or off-target binding forcompounds in the presence of whole blood (WB). The compound solutions inwhole blood and 1× kinetic running buffer (1×kb) were incubated at atemperature of 37° C. for 48 hours. Kinetic measurements for theincubated compound samples were performed with an Octet RED96 equippedwith streptavidin (SA) biosensor tips (ForteBio, Menlo Park, Calif.) at27° C. The ratio of the on-rates/binding signals in buffer and wholeblood were reported. A ratio <2 was considered to show no interference.

Results

The results are shown in Table 22.

TABLE 22 Parameter Compound A Compound E Ration of binding signal inwhole 1.5 1.4 blood/kinctic buffer to hu TNFa Ratio of binding signal inwhole 1.8 1.3 blood/kinetic buffer

Example 23: Summary of Tested Parameters

A summary of the parameter data for certain compounds is shown in Table23 below.

TABLE 23 Parameter Compound A Compound E Percent Monomer after two-  99.4% 99% step purification process Mass Spectrometry Profile IntactIntact pI as determined by IEF ~8.8 ~8.8 (heterogeneity) ThermalStability 57.9, 72.1, 82.9 67.6, 83.1 (differential scanningcalorimetry) Solubility 48 mg/ml 51 mg/ml Valence at pH 5.0 20.9 24.4Predicted Immunogenicity −37.69, −35.57 −31.1, −46.8 (EpiVax Score)Whole Blood Stability Maintained Maintained (human WB, 48 hrs at 37 C.);maintenance of binding to IL23 and TNFa

SEQUENCES SEQ ID NO Sequence 1EVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGLEWVSAITWNSGHIDYADSVEGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS 2DIQMTQSPSSLSASVGDRVTITCRASQGIRNYLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQRYNRAPYTFGQGTKVEIK 3QVQLVESOGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGNGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDV WGQGTTVTVSS 4EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIK 5QVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGDGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDV WGQGTTVTVSS 6EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIK 7QVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTV SS 8DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIK 9 GGGSGGG 10 LGGGSG 11FNRGES 12 VEPKSS 13DIQMTQSPSSLSASVGDRVTITCRASQGIRNYLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQRYNRAPYTFGQGTKVEIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSLGGGSGASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 14DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGEVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGLEWVSAITWNSGHIDYADSVEGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAKVSYLSTASSLDYWGQGTLVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 15DIQMTQSPSSLSASVGDRVTITCRASQGIRNYLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQRYNRAPYTFGQGTKVEIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSFNRGESASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 16DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGEVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGLEWVSAITWNSGHIDYADSVEGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAKVSYLSTASSLDYWGQGTLVTVSSVEPKSSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 17DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGEVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGLEWVSAITWNSGHIDYADSVEGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAKVSYLSTASSLDYWGQGTLVTVSSLGGGSGASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSIIEDPEVKFNWYVDGVEVIINAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 18DIQMTQSPSSLSASVGDRVTITCRASQGIRNYLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQRYNRAPYTFGQGTKVEIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 19DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGEVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGLEWVSAITWNSGHIDYADSVEGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAKVSYLSTASSLDYWGQGTLVTVSSFNRGESASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 20DIQMTQSPSSLSASVGDRVTITCRASQGIRNYLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQRYNRAPYTFGQGTKVEIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSVEPKSSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 21DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMIIWVRQAPGNGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSLGGGSGASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSIIEDPEVKFNWYVDGVEVIINAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 22EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 23DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGNGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSFNRGESASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRELQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 24EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSVEPKSSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 25EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSLGGGSGASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 26DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMIIWVRQAPGNGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 27EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSFNRGESASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLIIQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 28DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGNGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSVEPKSSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 29EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSLGGGSGASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 30DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGDGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 31DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGDGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSLGGGSGASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 32EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSLDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 33EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSFNRGESASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 34DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGDGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSVLPKSSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 35DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTEGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMIIWVRQAPGDGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSFNRGESASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSIIEDPEVKFNWYVDGVEVIINAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 36EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSVEPKSSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 37ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 38ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 39ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 40 EPKSCDKTHTCPPCP 41RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQLESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 42ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLIIQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 43ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 44DIQMTQSPSSLSASVGDRVTITCRASQGIRNYLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQRYNRAPYTFGQGTKVEIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSLGGGSGASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 45DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGEVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGLEWVSAITWNSGHIDYADSVEGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAKVSYLSTASSLDYWGQGTLVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 46DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGEVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGLEWVSAITWNSGHIDYADSVEGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAKVSYLSTASSLDYWGQGTLVTVSSFNRGESASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLIIQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 47DIQMTQSPSSLSASVGDRVTITCRASQGIRNYLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQRYNRAPYTFGQGTKVEIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSVEPKSSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 48DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGNGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSLGGGSGASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLEPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 49EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSLGGGSGRTVAAPSVFIEPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 50DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGNGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSFNRGESASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLIIQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 51EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSVEPKSSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSENRGEC 52EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSLGGGSGASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 53DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGNGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 54EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSLGGGSGASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSQEDPEVQFNWYVDGVEVIINAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 55DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGNGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 56EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSLGGGSGASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 57DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGNGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 58EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSLGGGSGASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 59DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGNGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 60EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTTSSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSFNRGESASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 61DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGNGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSVEPKSSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 62EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSFNRGESASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 63DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPLDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMIIWVRQAPGNGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSVEPKSSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 64EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSFNRGESASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 65DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGNGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSVEPKSSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 66EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSFNRGESASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 67DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGNGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSVEPKSSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 68DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGNGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSLGGGSGASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSIIEDPEVKFNWYVDGVEVIINAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 69EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 70DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGNGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSLGGGSGASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 71EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 72DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGNGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSLGGGSGASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 73EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 74DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGNGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSFNRGESASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 75EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSVEPKSSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 76DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGNGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSFNRGESASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 77EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIIIWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSVEPKSSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 78DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGNGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSFNRGESASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLIIQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 79EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSVEPKSSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 80EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSLGGGSGASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 81DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGDGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 82EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGTPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSLGGGSGASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 83DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRIITGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGDGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 84EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSLGGGSGASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 85DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGDGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 86EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIIIWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSLGGGSGASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSQEDPEVQFNWYVDGVEVIINAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 87DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGDGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 88EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSFNRGESASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 89DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGDGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSVEPKSSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 90EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSFNRGESASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNIIKPSNTKVDKRVEPKSCDKTIITCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMIIEALIINIIYTQKSLSLSPG 91DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGDGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSVEPKSSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 92EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSFNRGESASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 93DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGDGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSVEPKSSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 94EIVLTQSPATLSLSPGLRATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSFNRGESASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 95DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMIIWVRQAPGDGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSVEPKSSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 96DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGDGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSLGGGSGASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLIIQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 97EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 98DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTTSSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGDGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRALDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSLGGGSGASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 99EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKIIKVYACEVTIIQGLSSPVTKSENRGEC 100DIQMTQSPSSLSASVGDRVTITCKASRDVATAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGDGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSLGGGSGASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVIINAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 101EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 102DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGDGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSLGGGSGASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 103EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 104DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMIIWVRQAPGDGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSFNRGESASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 105EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSVEPKSSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 106DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGDGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSFNRGESASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 107EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSVEPKSSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 108DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCIIQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGDGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSFNRGESASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVIITFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDIIKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 109EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIIIWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSVEPKSSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGLC 110DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGDGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSFNRGESASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 111EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSVEPKSSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 112EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSLGGGSGASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVIITFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 113DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGNGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKIIKVYACEVTIIQGLSSPVTKSFNRGEC 114DIQMTQSPSSLSASVGDRVTITCRASQGIRNYLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQRYNRAPYTFGQGTKVEIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSLGGGSGASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTTSKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 115DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGEVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGLEWVSAITWNSGHIDYADSVEGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAKVSYLSTASSLDYWGQGTLVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 116DIQMTQSPSSLSASVGDRVTITCRASQGIRNYLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQRYNRAPYTFGQGTKVEIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSLGGGSGASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLIIQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 117DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGEVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGLEWVSAITWNSGHIDYADSVEGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAKVSYLSTASSLDYWGQGTLVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 118DIQMTQSPSSLSASVGDRVTITCRASQGIRNYLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQRYNRAPYTFGQGTKVEIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSLGGGSGASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 119DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGEVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGLEWVSAITWNSGHIDYADSVEGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAKVSYLSTASSLDYWGQGTLVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 120DIQMTQSPSSLSASVGDRVTITCRASQGIRNYLAWYQQKPGKAPKLLTYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQRYNRAPYTFGQGTKVEIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSFNRGESASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 121DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRIITGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGEVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGLEWVSAITWNSGHIDYADSVEGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAKVSYLSTASSLDYWGQGTLVTVSSVEPKSSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 122DIQMTQSPSSLSASVGDRVTITCRASQGIRNYLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQRYNRAPYTFGQGTKVEIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSFNRGESASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNIIKPSNTKVDKRVEPKSCDKTIITCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMIIEALIINIIYTQKSLSLSPG 123DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGEVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGLEWVSAITWNSGHIDYADSVEGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAKVSYLSTASSLDYWGQGTLVTVSSVEPKSSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 124DIQMTQSPSSLSASVGDRVTITCRASQGIRNYLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQRYNRAPYTFGQGTKVEIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSFNRGESASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVIINAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 125DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGEVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGLEWVSAITWNSGHIDYADSVEGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAKVSYLSTASSLDYWGQGTLVTVSSVEPKSSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 126DIQMTQSPSSLSASVGDRVTITCRASQGIRNYLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQRYNRAPYTFGQGTKVEIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSFNRGESASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 127DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGEVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGLEWVSAITWNSGHIDYADSVEGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAKVSYLSTASSLDYWGQGTLVTVSSVEPKSSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 128DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGEVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGLEWVSAITWNSGHIDYADSVEGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAKVSYLSTASSLDYWGQGTLVTVSSLGGGSGASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 129DIQMTQSPSSLSASVGDRVTITCRASQGIRNYLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQRYNRAPYTFGQGTKVEIKGGGSGGGGQVQLVQSGAELVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 130DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRIITGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGEVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGLEWVSAITWNSGHIDYADSVEGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAKVSYLSTASSLDYWGQGTLVTVSSLGGGSGASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVIITFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 131DIQMTQSPSSLSASVGDRVTITCRASQGIRNYLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQRYNRAPYTFGQGTKVEIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKIIKVYACEVTIIQGLSSPVTKSFNRGEC 132DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGEVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGLEWVSAITWNSGHIDYADSVEGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAKVSYLSTASSLDYWGQGTLVTVSSLGGGSGASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 133DIQMTQSPSSLSASVGDRVTITCRASQGIRNYLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQREDVATYYCQRYNRAPYTFQGTKVEIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIIIWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 134DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGEVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGLEWVSAITWNSGHIDYADSVEGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAKVSYLSTASSLDYWGQGTLVTVSSLGGGSGASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 135DIQMTQSPSSLSASVGDRVTITCRASQGIRNYLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQRYNRAPYTFGQGTKVEIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 136EIVLTQSPATLSLSPGERATLSCRASQSVYSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPFTFGPGTKVDIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSLGGGSGASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 137DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYAMHWVRQAPGNGLEWVAFMSYDGSNKKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGIAAGGNYYYYGMDVWGQGTTVTVSSLGGGSGRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 138DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGEVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGLEWVSAITWNSGHIDYADSVEGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAKVSYLSTASSLDYWGQGTLVTVSSFNRGESASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSIIEDPEVKFNWYVDGVEVIINAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 139DIQMTQSPSSLSASVGDRVTITCRASQGIRNYLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQRYNRAPYTFGQGTKVEIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSVEPKSSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 140DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGEVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGLEWVSAITWNSGHIDYADSVEGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAKVSYLSTASSLDYWGQGTLVTVSSFNRGESASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 141DIQMTQSPSSLSASVGDRVTITCRASQGIRNYLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQRYNRAPYTFGQGTKVEIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSVEPKSSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 142DIQMTQSPSSLSASVGDRVTITCKASRDVAIAVAWYQQKPGKVPKLLIYWASTRHTGVPSRFSGSGSRTDFTLTISSLQPEDVADYFCHQYSSYPFTFGSGTKLEIKGGGSGGGGEVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMIIWVRQAPGKGLEWVSAITWNSGIIIDYADSVEGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAKVSYLSTASSLDYWGQGTLVTVSSFNRGESASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAREFLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSQEDPEVQFNWYVDGVEVIINAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 143DIQMTQSPSSLSASVGDRVTITCRASQGIRNYLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQRYNRAPYTFGQGTKVEIKGGGSGGGGQVQLVQSGAEVKKPGSSVKVSCKASGYTFTDQTIHWMRQAPGQGLEWIGYIYPRDDSPKYNENFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCAIPDRSGYAWFIYWGQGTLVTVSSVEPKSSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 144MSTESMIRDVELAEEALPKKTGGPQGSRRCLFLSLFSFLIVAGATTLFCLLHFGVIGPQREEFPRDLSLISPLAQAVRSSSRTPSDKPVAHVVANPQAEGQLQWLNRRANALLANGVELRDNQLVVPSEGLYLIYSQVLFKGQGCPSTHVLLTHTISRIAVSYQTKVNLLSAIKSPCQRETPEGAEAKPWYEPIYLGGVFQLEKGDRLSAEINRPDYLDFAESGQVYFGIIAL 145MLGSRAVMLLLLLPWTAQGRAVPGGSSPAWTQCQQLSQKLCTLAWSAHPLVGHMDLREEGDEETTNDVPHIQCGDGCDPQGLRDNSQFCLQRIHQGLIFYEKLLGSDIFTGEPSLLPDSPVGQLHASLLGLSQLLQPEGHHWETQQIPSLSPSQPWQRLLLRFKILRSLQAFVAVAARV FAHGAATLSP146 DYAMH 147 AITWNSGHIDYADSVEG 148 VSYLSTASSLDY 149 RASQGIRNYLA 150AASTLQS 151 QRYNRAPYT 152 SYAMH 153 FMSYDGSNKKYADSVKG 154 NYYYYGMDV 155RASQSVYSYLA 156 DASNRAT 157 QQRSNWPPFT 158 DQTIH 159 YIYPRDDSPKYNENFKG160 PDRSGYAWFIY 161 KASRDVAIAVA 162 WASTRHT 163 HQYSSYPFT

Other Embodiments

All of the features disclosed in this specification may be combined inany combination. Each feature disclosed in this specification may bereplaced by an alternative feature serving the same, equivalent, orsimilar purpose. Thus, unless expressly stated otherwise, each featuredisclosed is only an example of a generic series of equivalent orsimilar features.

From the above description, one skilled in the art can easily ascertainthe essential characteristics of the present disclosure, and withoutdeparting from the spirit and scope thereof, can make various changesand modifications of the disclosure to adapt it to various usages andconditions. Thus, other embodiments are also within the claims.

What is claimed is:
 1. A method of treating an autoimmune orinflammatory disease comprising administering a therapeuticallyeffective amount of a compound comprising a first polypeptide and asecond polypeptide, wherein: (A) said first polypeptide comprises: (i) alight chain variable domain of a first immunoglobulin (VL1) specific fora first target protein; (ii) a heavy chain variable domain of a secondimmunoglobulin (VH2) specific for a second target protein; and (iii) ahinge region, a heavy chain constant region 2 (CH2) and a heavy chainconstant region 3 (CH3); and (B) said second polypeptide comprises: (i)a light chain variable domain of the second immunoglobulin (VL2)specific for said second target protein; (ii) a heavy chain variabledomain of the first immunoglobulin (VH1) specific for said first targetprotein; (C) in said first and second polypeptides of (A) and (B): (i)said VL1 and VH1 associate to form a binding site that binds said firsttarget protein; (ii) said VL2 and VH2 associate to form a binding sitethat binds said second target protein; (iii) said heavy chain constantregion 2 (CH2) comprises a tyrosine at position 252, a threonine atposition 254 and a glutamic acid a position 256, numbered according tothe EU index as in Kabat for the CH2 of a human IgG antibody; and (iv)said first target protein is TNF-alpha and said second target protein isIL-23A or said first target protein is IL-23A and said second targetprotein is TNF-alpha, and (D) in said first and second polypeptides of(A), (B) and (C): (i) said VL1 comprises the amino acid sequence of SEQID NO:2, said VH1 comprises the amino acid sequence of SEQ ID NO:1, saidVL2 comprises the amino acid sequence of SEQ ID NO:8 and said VH2comprises the amino acid sequence of SEQ ID NO:7; or (ii) said VL1comprises the amino acid sequence of SEQ ID NO:4 or 6, said VH1comprises the amino acid sequence of SEQ ID NO:3 or 5, said VL2comprises the amino acid sequence of SEQ ID NO:8 and said VH2 comprisesthe amino acid sequence of SEQ ID NO:7; or (iii) said VL1 comprises theamino acid sequence of SEQ ID NO:8, said VH1 comprises the amino acidsequence of SEQ ID NO:7, said VL2 comprises the amino acid sequence ofSEQ ID NO:2 and said VH2 comprises the amino acid sequence of SEQ IDNO:1; or (iv) said VL1 comprises the amino acid sequence of SEQ ID NO:8,said VH1 comprises the amino acid sequence of SEQ ID NO:7, said VL2comprises the amino acid sequence of SEQ ID NO:4 or 6 and said VH2comprises the amino acid sequence of SEQ ID NO:3 or
 5. 2. The method ofclaim 1, wherein in (D)(ii) said VL1 comprises the amino acid sequenceof SEQ ID NO:4, said VH1 comprises the amino acid sequence of SEQ IDNO:3, said VL2 comprises the amino acid sequence of SEQ ID NO:8 and saidVH2 comprises the amino acid sequence of SEQ ID NO:7.
 3. The method ofclaim 1, wherein in (D)(ii) said VL1 comprises the amino acid sequenceof SEQ ID NO:6, said VH1 comprises the amino acid sequence of SEQ IDNO:5, said VL2 comprises the amino acid sequence of SEQ ID NO:8 and saidVH2 comprises the amino acid sequence of SEQ ID NO:7.
 4. The method ofclaim 1, wherein in (D)(iv) said VL2 comprises the amino acid sequenceof SEQ ID NO:4, said VH2 comprises the amino acid sequence of SEQ IDNO:3, said VL1 comprises the amino acid sequence of SEQ ID NO:8 and saidVH1 comprises the amino acid sequence of SEQ ID NO:7.
 5. The method ofclaim 1, wherein in (D)(iv) said VL2 comprises the amino acid sequenceof SEQ ID NO:6, said VH2 comprises the amino acid sequence of SEQ IDNO:5, said VL1 comprises the amino acid sequence of SEQ ID NO:8 and saidVH1 comprises the amino acid sequence of SEQ ID NO:7.
 6. The method ofclaim 1, wherein said first polypeptide further comprises a first linkerbetween said VL1 and said VH2 and said second polypeptide furthercomprises a second linker between said VL2 and said VH1.
 7. The methodof claim 6, wherein said first linker or said second linker comprisesthe amino acid sequence of GGGSGGG (SEQ ID NO:9).
 8. The method of claim6, wherein said first linker and said second linker comprise the aminoacid sequence of GGGSGGG (SEQ ID NO:9).
 9. The method of claim 1,wherein said first polypeptide further comprises a heavy chain constantregion 1 domain (CH1) and said second polypeptide further comprises alight chain constant region domain (CL), wherein said CL and said CH1are associated together via a disulfide bond to form a Cl domain. 10.The method of claim 9, wherein said first polypeptide further comprisesa third linker between said VH2 and said CH1 and said second polypeptidefurther comprises a fourth linker between said VH1 and said CL.
 11. Themethod of claim 10, wherein said third linker comprises the amino acidsequence of FNRGES (SEQ ID NO:11).
 12. The method of claim 10, whereinsaid fourth linker comprises the amino acid sequence of VEPKSS (SEQ IDNO:12).
 13. The method of claim 10, wherein said third linker comprisesthe amino acid sequence of FNRGES (SEQ ID NO:11) and said fourth linkercomprises the amino acid sequence of VEPKSS (SEQ ID NO:12).
 14. Themethod of claim 10, wherein said third linker or said fourth linkercomprises the amino acid sequence of LGGGSG (SEQ ID NO:10).
 15. Themethod of claim 10, wherein said third linker and said fourth linkercomprise the amino acid sequence of LGGGSG (SEQ ID NO:10).
 16. Themethod of claim 1, wherein said heavy chain constant region 2 (CH2) of(C)(iii) comprises an alanine at positions 234 and an alanine atposition 235, numbered according to the EU index as in Kabat for the CH2of a human IgG antibody.
 17. The method of claim 1, wherein the aminoacid sequence of said hinge region, said heavy chain constant region 2(CH2) or said heavy chain constant region 3 (CH3) is derived from a IgG1or from a IgG4.
 18. The method of claim 1, wherein said hinge regioncomprises the amino acid sequence of EPKSCDKTHTCPPCP (SEQ ID NO:40). 19.The method of claim 1, wherein said compound comprises two said firstpolypeptides and two said second polypeptides, wherein said two firstpolypeptides are associated together via at least one disulfide bond.20. The method of claim 1, wherein: (i) said first polypeptide comprisesthe amino acid sequence of SEQ ID NO:13 and said second polypeptidecomprises the amino acid sequence of SEQ ID NO:14; (ii) said firstpolypeptide comprises the amino acid sequence of SEQ ID NO:15 and saidsecond polypeptide comprises the amino acid sequence of SEQ ID NO:16;(iii) said first polypeptide comprises the amino acid sequence of SEQ IDNO:17 and said second polypeptide comprises the amino acid sequence ofSEQ ID NO:18; (iv) said first polypeptide comprises the amino acidsequence of SEQ ID NO:19 and said second polypeptide comprises the aminoacid sequence of SEQ ID NO:20; (v) said first polypeptide comprises theamino acid sequence of SEQ ID NO:21 and said second polypeptidecomprises the amino acid sequence of SEQ ID NO:22; (vi) said firstpolypeptide comprises the amino acid sequence of SEQ ID NO:23 and saidsecond polypeptide comprises the amino acid sequence of SEQ ID NO:24;(vii) said first polypeptide comprises the amino acid sequence of SEQ IDNO:25 and said second polypeptide comprises the amino acid sequence ofSEQ ID NO:26; (viii) said first polypeptide comprises the amino acidsequence of SEQ ID NO:27 and said second polypeptide comprises the aminoacid sequence of SEQ ID NO:28; (ix) said first polypeptide comprises theamino acid sequence of SEQ ID NO:29 and said second polypeptidecomprises the amino acid sequence of SEQ ID NO:30; (x) said firstpolypeptide comprises the amino acid sequence of SEQ ID NO:31 and saidsecond polypeptide comprises the amino acid sequence of SEQ ID NO:32;(xi) said first polypeptide comprises the amino acid sequence of SEQ IDNO:33 and said second polypeptide comprises the amino acid sequence ofSEQ ID NO:34; or (xii) said first polypeptide comprises the amino acidsequence of SEQ ID NO:35 and said second polypeptide comprises the aminoacid sequence of SEQ ID NO:36.
 21. The method of claim 20, wherein saidcompound comprises two said first polypeptides and two said secondpolypeptides, wherein said two first polypeptides are associatedtogether via at least one disulfide bond and wherein each of said firstpolypeptide is associated to one said second polypeptide via at leastone disulfide bond.
 22. The method of claim 20, wherein said compoundcomprises two said first polypeptides and two said second polypeptides,wherein each of said first polypeptides comprises a CH1, a CH2 and a CH3and each of said second polypeptides comprises a CL and wherein the CH2and CH3 of one of the first polypeptides associates with the CH2 and CH3of the other of the first polypeptides and the CH1 of each said firstpolypeptides associates with the CL of one said second polypeptides toform a tetravalent molecule.
 23. The method of claim 1, wherein saidautoimmune disease is selected from the group consisting of rheumatoidarthritis, psoriasis, type 1 diabetes, systemic lupus erythematosus,transplant rejection, autoimmune thyroid disease (Hashimoto's disease),sarcoidosis, scleroderma, granulomatous vasculitis, Crohn's disease,ulcerative colitis, Sjogren's disease, ankylosing spondylitis, psoriaticarthritis, polymyositis dermatomyositis, polyarteritis nodosa,immunologically mediated blistering skin diseases, Behcet's syndrome,multiple sclerosis, systemic sclerosis, Goodpasture's disease and immunemediated glomerulonephritis.
 24. The method of claim 1, wherein saidinflammatory disease is selected from the group consisting of rheumatoidarthritis, systemic lupus erythematosus, alopecia areata, ankylosingspondylitis, antiphospholipid syndrome, autoimmune Addison's disease,autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune inner eardisease, autoimmune lymphoproliferative syndrome (ALPS), autoimmunethrombocytopenic purpura (ATP), Behcet's disease, bullous pemphigoid,cardiomyopathy, celiac sprue-dermatitis, chronic fatigue syndrome immunedeficiency syndrome (CFIDS), chronic inflammatory demyelinatingpolyneuropathy, cicatricial pemphigoid, cold agglutinin disease, Crestsyndrome, Crohn's disease, Dego's disease, dermatomyasitis,dermatomyositis, juvenile, discoid lupus, essential mixedcryoglobulinemia, fibromyalgia, fibromyositis, Grave's disease,Guillain-Barre, Hashimoto's thyroiditis, idiopathic pulmonary fibrosis,idiopathic thrombocytopenia purpura (ITP), IgA nephropathy, insulindependent diabetes (Type I), juvenile arthritis, Meniere's disease,mixed connective tissue disease, multiple sclerosis, myasthenia gravis,pemphigus vulgaris, pernicious anemia, polyarteritis nodosa,polychondritis, polyglancular syndromes, polymyalgia rheumatica,polymyositis, dermatomyositis, primary agammaglobulinemia, primarybiliary cirrhosis, psoriasis, Raynaud's phenomenon, Reiter's syndrome,rheumatic fever, sarcoidosis, scleroderma, Sjogren's syndrome, stiff-mansyndrome, Takayasu arteritis, temporal arteritis/giant cell arteritis,ulcerative colitis, uveitis, vasculitis, vitiligo, and Wegener'sgranulomatosis.
 25. The method of claim 1, wherein said autoimmune orinflammatory disease is selected from the group consisting of Crohn'sdisease, ankylosing spondylitis, or psoriatic arthritis.